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 ArticleISSN: 0974-6943

Available online throughwww.jpronline.info

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

INTRODUCTIONThe 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 56species of herbaceous plant in sunflower family. Tagetes genus is originatedin North and South America. Now-a-days it is also cultivated in Asiancountries like India, Bhutan, Nepal, China [2].Use of Tagetes in magical,religious and medicinal uses have been mentioned in history. Tagetes is usedin the treatment of hiccups [3]. T.erecta is used in garlands to decorate housesand Idols of God during the festivals in India. Since ancient time parts of thisplant has been used for medicinal purposes and for the skin wash and yellowdye is used as by Cherokee [4]. Marigold is commonly used in food additivesas a coloring agent and as animal food in fodder (dried flower meal & extractused as supplement for poultry feed). It is used as an ornamental plant andalso in tannin or dye industry [5]. It is also used in medicine (folklore) and asa poison for non vertebrates and in plant pest control [6]. Marigold has socialvalue because of its use in religious purposes. The Tagetes is used to adornIdols during festivals in India because of its stability [7]. Tagetes erecta isused as a food color in the African countries because of its richness incarotenoid leutin [8]. The species Tagetes lucida is used to prepare sweetmedicinal tea [9]. Tagetes minuta is used to make a popular potato dish calledOcopa [10]. The essential oil of Tagetes is extracted from its leaves, stems,stalk and flowers by steam distillation [11]. The main components of this oilare limonene, ocimene, lagetone and valeric acid [12]. Essential oil of Tagetescan 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 inwound burns, skin diseases, athlete’s foot, colitis and dermatitis [14]. Fromscientific 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

ABSTRACTThe antibacterial effect of Mexican marigold (Tagetes erecta) leaf extract at room temperature against 10 gram positive multidrug resistant bacterialisolates including Enterococcus faecalis, Enterococcus faecium. Staphylococcus epidermidis, Staphylococcus aureus, Bacillus cereus, Erysipelothrixrhusiopathiae, Propionibacterium acne, Streptococcus pneumoniae, Streptococcus agalactine, Staphylococcus saprophyticus and 6 gram negative multidrugresistant 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-organismwas obtained for Acinetobacter baumannii (Acitvity Index = 0.913333333) and Propionibacterium acne (Acitvity Index = 0.906666667) and minimumwas for Streptococcus pneumonia (Activity Index = 0.026666667). The results suggest that species of Mexican marigold i.e. Tagetes erecta hasantibacterial effect against airborne disease causing gram positive and gram negative bacteria and mainly against skin infection causing bacteria, and hencecan 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 Tagetesspecies and they have been used against gram negative and gram positivebacteria in vitro [15]. Tagetes erecta is also used as interplant with cowpeafor control of nematode pests [16]. Tagetes erecta essential oil also hasantioxidant properties [17].

MATERIALS AND METHODS

Microorganisms usedA total of 10 gram positive and 3 gram negative organisms were used in thestudy. The isolates of Enterococcus faecalis, Enterococcus faecium.Staphylococcus epidermidis, Staphylococcus aureus, Bacillus cereus,Erysipelothrix rhusiopathiae, Propionibacterium acne, Streptococcuspneumoniae, Streptococcus agalactine, Staphylococcus saprophyticus,Escherichia coli, Klebsiella pneumoniae, Pseudomonas auregenosa,Salmonella enteriditis, Acinetobacter baumannii and Alcaligen faecalis wereobtained from the culture collection centre of Institute of MicrobialTechnology (IMTECH), Chandigarh, India.

Media UsedThe media used in the present investigation was Nutrient Agar media, whichwas obtained from Hi – Media Laboratories Ltd., Mumbai, India.

Antibiotic Gentamycin usedThe antibiotic standard used in this investigation was Gentamicin G30susceptibility test discs (30 mg per disc) which were obtained from Hi –Media Laboratories Ltd., Mumbai, India.

Aqueous stock preparations of Mexican marigold leafFresh Mexican marigold (Tagetes erecta) leaves were collected from nurseryat VIT University, Tamil Nadu, India. 50 gm of surface sterilised Mexicanmarigold leaves were gently grinded using mortar and pestle. The crudeextract 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 leafextract which was separated in conical flask. The supernatant extracted wasused as stock for antibacterial activity and stored in the refrigerator.

Re – culturing of micro organisms used or Enrichment of cultureThe bacterial cultures were maintained in Peptone Saline Water.

Analyzing Antibacterial Activity of Mexican marigold (Tagetes erecta)leaf extract using Agar well diffusion assay100µl of each bacterial strain was used to make lawn culture on sterileNutrient 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. Thereshould be sufficient distance between the wells and petriplate wall to avoidoverlapping of zone of inhibition. The plates were incubated in an uprightposition at 37°C for 24 hours. The diameter of zone of inhibition wasmeasured in millimeter and the results were recorded. The inhibition zoneswith diameter less than 8mm were not considered as having antibacterialactivity.

Antibiotic sensitivity testingThe test microorganisms were also tested for their sensitivity against theantibiotic (Gentamicin 30 mg). Using sterile cotton swabs, the enrichedcultures were aseptically swabbed on the surface of sterile nutrient agarplates. Using ethanol dipped and flamed forceps, the antibiotic (Gentami-cin, 30 mg) disc was aseptically placed over the agar plates sufficientlyseparated 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 IndexFollowing 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 similarlyzone of inhibition of Antibiotic (Gentamicin) were measured.

RESULT AND DISCUSSION

Activity Index of Mexican marigold (Tagetes erecta) leaf extractFrom the triplicate readings average zone of inhibition for Mexican mari-gold leaf extract, average zone of inhibition for Gentamicin (30 mg) andusing 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 foundin Acinetobacter baumannii (Acitvity Index = 0.913333333) and minimumfor Streptococcus pneumonia (Activity Index = 0.026666667). Also Mexi-can marigold leaf has very less antibacterial effect on Streptococcusagalactine, Staphylococcus saprophyticus having activity index 0.037533333and 0.047621428 respectively. For other gram positive bacterial strainsMexican marigold has activity index between 0.763 and 9.066.

For gram negative bacterial strains maximum antibacterial effect was foundin 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 DISCUSSIONThe activity index of Mexican marigold for different gram positive and gramnegative isolated bacterial strain were calculated after antibacterial testingusing agar well diffusion method and antibacterial sensitivity testing withGentamicin. The maximum antibacterial effect of Mexican marigold leafextract among those micro-organism was obtained for Acinetobacterbaumannii (Acitvity Index = 0.913333333) and Propionibacterium acne(Acitvity Index = 0.906666667) and minimum was for Streptococcuspneumoniae (Activity Index = 0.026666667). Reason for minimum activityindex on Streptococcus pneumoniae is its capsulated structure. The resultssuggest us that species of marigold i.e. Tagetes erecta has antibacterialeffect against airborne disease causing gram positive and gram negativebacteria and mainly against skin infection causing bacteria, so it can beuseful in developing drugs for diseases like dermatitis, acne, skin races andalso 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 AverageNo bacterial species of inhibition of inhibition Activity

of Tagetes erecta of Gentamicin Indexleaf extract (in mm)(in mm)

1. Enterococcus faecalis 1.123333 1.466666667 0.7633333332. Enterococcus faecium 1.156667 1.466666667 0.7833333333. Staphylococcus epidermidis 0.893333 1.143333333 0.7766666674. Staphylococcus aureus 1.166667 1.466666667 0.795. Bacillus cereus 1.15 1.46 0.7733333336. Erysipelothrix rhusiopathiae 1.196667 1.476666667 0.8066666677. Propionibacterium acne 1.34 1.46 0.9066666678. Streptococcus pneumoniae 0.006667 0.24 0.0266666679. Streptococcus agalactine 0.006667 0.25. 0.03753333310. Staphylococcus saprophyticus 0.006667 0.14 0.04762142811. Escherichia coli 1.256667 1.49 0.81666666712. Klebsiella pneumoniae 0.403333 0.893333333 0.44333333313. Pseudomonas auregenosa 1.23 1.406666667 0.8714. Salmonella enteriditis 0.306667 0.666666667 0.45333333315. Acinetobacter baumannii 1.196667 1.3 0.91333333316. Alcaligen faecalis 0.983333 1.166666667 0.84

Figure 1: Graphical analysis of Activity Index of various isolated gram positivebacterial strain.

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

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

Figure 3: Zone of inhibition by mexicanmarigold on Erysipelothrix rhusiopathiae.

Figure 4: Zone of inhibition by mexicanmarigold on Propionibacterium acne.

Figure 5: Zone of inhibi t ion bymexican marigold on Escherichia coli.

Figure 6: Zone of inhibi t ion bymexican marigold on Pseudomonasauregenosa.

Figure 7: Zone of inhibition byMexican marigold on Acinetobacterbaumannii.

Figure 9: Zone of inhibi t ion byMexican marigold on Alcal igenfaecalis.

Figure 10: Zone of inhibition byMexican marigold on Streptococcuspneumonia, zero activity index.

Figure 11: Zone of inhibition byMexican marigold on Streptococcusagalactine, zero activity index.

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