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A Comparative study of Antimicrobial activity of Datura
(Leaf, Seed), Neem (Leaf, Bark) and Orange (Peel, waste) on
Escherichia coli and Paenibacillus barcinonesis
Vivek Mishra1, Deepesh Kumar
2, V.K.Srivastava
3, Ashish shukla
4,
Aarti Gautam5, Shivani Shukla
6 , Richa Saxena
7, Abhishek Upadhyay
8,
Saksham khusharia9
1,2,3,4,5,7,8,9 Post Graduate Department of Biotechnology,
RBS Engineering Technical Campus, Bichpuri, Agra, AKTU Lucknow, (India)
6Department of Bioengineering, Integral University, Lucknow, India
ABSTRACT
In this present study was conducted to identify bacteria (Paenibacillus barcinonesis) by using biochemical test and
then to identify antimicrobial investigation against (leaf extract, bark extract) of Neem, (peel extract, waste extract)
of Orange and (leaf extract, seed extract) of Datura which is collected from the local market and Raja Balwant
singh Engineering Technical Campus Bichpuri, Agra (U.P.) India. Antimicrobial activity of crude extract
(Methanol, Isopropanol, Chloroform, Water, Butanol) were tested by using Well Diffusion Method against two
Bacterial strains (Escherichia coli and Paenibacillus barcinonesis). Neem leaves methanol crude extract produced
maximum zone of inhibition at 24 hrs, 48hrs and (2.26 ± 0.05 cm), (2.5 ± 0.17cm) and (2.6 ± 0.17cm) respectively
against Escherichia coli. Neem leaves water crude extract produced maximum zone of inhibition at 24hrs, 48hrs
and 72hrs (1.36 ± 1.01cm), (1.4 ± 0.1cm) and (1.5 ± 0.1cm) respectively against Paenibacillus barcinonesis.
Methanol crude extract Bark of Neem showed maximum zone of inhibition at 24hrs, 48hrs and 72hrs (1.5 ± 0.43cm),
(2.06 ± 0.05cm) and (2.13 ± 0.11cm) respectively against Escherichia coli. Methanol crude extract Bark of Neem
showed maximum zone of inhibition at 24hrs, 48hrs and 72hrs (1.3 ± 0cm), (1.4 ± 0.05cm) and (1.36 ± 0.05cm)
respectively against Paenibacillus barcinonesis. Orange peel methanol crude extract and butanol crude extract
produced maximum zone of inhibition at 24hrs and 48hrs, 72hrs (1.4 ± 0.05cm) and (1.33 ± 0.11cm), (1.46 ±
0.15cm) respectively against Escherichia coli. Orange peel methanol crude extract and water crude extract
produced maximum zone of inhibition at 24hrs and 48hrs, 72hrs (0.86 ± 0.05cm) and (0.83 ± 0.15cm), (0.86 ±
0.05cm) against Paenibacillus barcinonesis. Methanol and Butanol crude extract Waste of Orange showed
ma++++++ximum zone of inhibition at 24hrs and 48hrs, 72hrs (1.6 ± 0.51cm) and (2.7 ± 0cm), (1.03 ± 0.05cm)
against Escherichia coli. Chloroform crude extract Waste of Orange showed maximum zone of inhibition at 24hrs,
48hrs and 72hrs (0.9 ± 0.1cm), (0.9 ± 0.1cm) and (0.93 ± 0.05cm) against Paenibacillus barcinonesis. Methanol
and Water crude extract leaves of Datura showed maximum zone of inhibition at 24hrs, 72hrs and 48hrs (1.36 ±
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0.05cm), (1.56 ± 0.05cm) and (1.46 ± 0.05cm) against Escherichia coli. Methanol and Butanol crude extract leaves
of Datura showed maximum zone of inhibition at 24hrs, 48hrs and 72hrs (3.6 ± 0cm), (3.6 ± 0.1cm) and (3.8 ±
0.05cm) against Paenibacillus barcinonesis. Datura seed methanol crude extract crude extract produced maximum
zone of inhibition at 24hrs, 48hrs and 72hrs (1.06 ± 0.28cm), (1.5 ± 0.05cm), and (1.56 ± 0.05cm) respectively
against Escherichia coli. Datura seed methanol crude extract crude extract produced maximum zone of inhibition at
24hrs, 48hrs and 72hrs (1.33 ± 0.05cm), (1.53 ± 0.15cm), and (1.6 ± 0.2cm) respectively against Paenibacillus
barcinonesis.
Keywords- Datura,Escherichia coli, Neem, Orange, Paenibacillus barcinonesis
I INTRODUCTION
Datura metel Linn (Thorn-apple, Devil trumpet, Solanaceae) is a Nigerian medicinal plant widely used in
phytomedicine to cure diseases such as asthma, cough, convulsion and insanity. The leaves and seeds are widely
used in herbal medicine as anesthetic, antispasmodic, bronchodilator and as hallucinogenic (Duke and Ayensu, 1985
and Dabur et al., 2004).). A variety of phytochemicals have been found to occur in D.metel. These phytoconstituents
comprises alkaloids, flavonoids, phenols, tannins, saponins and sterols.The solanaceous alkaloids hyoscyamine and
scopolamines have been isolated from D. metel (Chopra etal., 1986, Oliver-Bever, 1986 ).
Neem (Azadirachta indica) commonly called ‘India Lilac’ or ‘Margosa’, belongs to the family
Meliaceae, subfamily Meloideae and tribe Melieae. Neem is the most versatile, multifarious trees of tropics, with
immense potential. It possesses maximum useful non-wood products (leaves, bark, flowers, fruits, seed, gum, oil
and neem cake) than any other tree species. Various parts of the neem tree have been used as traditional Ayurvedic
medicine in India.
Neem oil and the bark and leaf extracts have been therapeutically used as folk medicine to control leprosy, intestinal
helminthiasis, respiratory disorders, and constipation and also as a general health promoter. Neem oil finds use to
control various skin infections. Bark, leaf, root,flower and fruit together cure blood morbidity, biliary afflictions,
itching, skin ulcers, burning sensations and phthisis.
Neem tree has adaptability to a wide range of climatic, topographic and edaphic factors. It thrives well in dry, stony
shallow soils and even on soils having hard calcareous or clay pan, at a shallow depth. Neem tree requires little
water and plenty of sunlight.
Peels are generally wasted while the citrus fruits are mainly used in juice processing industries. Very large amounts
of by-product are formed as wastes during the production of citrus juices. Citrus is one of the most important
commercial fruit crops grown in all continents of the world. Oranges and Lemon are an important medicinal plant of
the family Rutaceae. It is cultivated mainly for its alkaloids, which are having anticancer activities and the
antibacterial potential in crude extracts of different parts (viz., leaves, stem, root and flower) of Lemon against
clinically significant bacterial strains has been reported. Citrus fruits are mainly used by juice processing industries
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while the peels are generally wasted. Since the juice yield of citrus is less half of the fruit weight, very large amounts
of by product wastes, such as peels are formed every year.
II MATERIAL AND METHODS
Plants Materials
The Neem leaves & Bark, Datura Leaves & Seeds were collected from RBS ETC, Bichpuri, Agra(U.P.) India and
Orange Leaves and Waste was collected from local market Bichpuri, Agra.
Microorganisms
Paenibacillus barcinonesis was isolated agriculture soil sample and E.coli was provided by RBS ETC, Bichpuri,
Agra.
Reference Drug
Reference drug used in this study was Ampicillin (10µl/disc).
Extraction- All collected materials were shade dried at room temperature for about 5-7 days. Then the samples
were cut into small pieces and powdered in a Mortar and Pestle. Extractions were carried out with Methanol,
Isopropanol, Butanol, Chloroform and Water solvent of bioactive compounds. After 24hrs the extract was filtered
with whatman filter paper then crude extract found for antimicrobial investigation.
Testing for Antimicrobial
Different concentrations of methanol, isopropanol, butanol, chloroform and water extracts of Datura,Neem and
Orange (10%, 20% and 30%). Take 10g of plant powder was added 100ml methanol, isopropanol, butanol,
chloroform and water into a conical flask for 24hrs.after 24hrs the extract were filtrate then used.(Harbone JB,
1973).
III RESULTS
We take soil sample from agricultural land from the RBS ETC, Bichpuri, Agra (U.P.) India then dilution of this
sample 1/10th
rule. Take five test tube and then mark 100-10
-4 and the last test tube diluted sample mixed well and
take 100µl in the Nutrient Agar petri plate disc for 24hrs incubated. Then I focused a single colony that plate after
then steak on a Nutrient Agar plate further incubated for 24hrs.To identified bacteria we confirmed by biochemical
tests then analyzed by Abis online which is found Paenibacillus barcinonesis.
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Serial Dilution Single colony Isolation
Biochemical Tests
Table-1
Gram-Staning
+Ve
Mannitol Test
+Ve
Starch Hydrolysis
-Ve
Catalase Activity
+Ve
Gelatin Test
+Ve
Methyl-Red Test
+Ve
Voges-Proskers Test -Ve
H2S Test
+Ve
Citrate Test
-Ve
Urease Test
-Ve
Gram’s staining Catalase Test Citrate Test
Gelatin test H2S Test Mannitol Test
Methyl red Starch Test Vp Test
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Antimicrobial activity testing by well diffusion method the antibiotic Ampicillin take as a control and then
Antimicrobial activity of crude extract (Methanol, Isopropanol, Chloroform, Water, Butanol) were tested against
two Bacterial strains (Escherichia coli and Paenibacillus barcinonesis). Neem leaves methanol crude extract
produced maximum zone of inhibition at 24 hrs, 48hrs and (2.26 ± 0.05 cm), (2.5 ± 0.17cm) and (2.6 ± 0.17cm)
respectively against Escherichia coli. Neem leaves water crude extract produced maximum zone of inhibition at
24hrs, 48hrs and 72hrs (1.36 ± 1.01cm), (1.4 ± 0.1cm) and (1.5 ± 0.1cm) respectively against Paenibacillus
barcinonesis. Methanol crude extract Bark of Neem showed maximum zone of inhibition at 24hrs, 48hrs and 72hrs
(1.5 ± 0.43cm), (2.06 ± 0.05cm) and (2.13 ± 0.11cm) respectively against Escherichia coli. Methanol crude extract
Bark of Neem showed maximum zone of inhibition at 24hrs, 48hrs and 72hrs (1.3 ± 0cm), (1.4 ± 0.05cm) and (1.36
± 0.05cm) respectively against Paenibacillus barcinonesis. Orange peel methanol crude extract and butanol crude
extract produced maximum zone of inhibition at 24hrs and 48hrs, 72hrs (1.4 ± 0.05cm) and (1.33 ± 0.11cm), (1.46 ±
0.15cm) respectively against Escherichia coli. Orange peel methanol crude extract and water crude extract produced
maximum zone of inhibition at 24hrs and 48hrs, 72hrs (0.86 ± 0.05cm) and (0.83 ± 0.15cm), (0.86 ± 0.05cm)
against Paenibacillus barcinonesis. Methanol and Butanol crude extract Waste of Orange showed maximum zone of
inhibition at 24hrs and 48hrs, 72hrs (1.6 ± 0.51cm) and (2.7 ± 0cm), (1.03 ± 0.05cm) against Escherichia coli.
Chloroform crude extract Waste of Orange showed maximum zone of inhibition at 24hrs, 48hrs and 72hrs (0.9 ±
0.1cm), (0.9 ± 0.1cm) and (0.93 ± 0.05cm) against Paenibacillus barcinonesis. Methanol and Water crude extract
leaves of Datura showed maximum zone of inhibition at 24hrs, 72hrs and 48hrs (1.36 ± 0.05cm), (1.56 ± 0.05cm)
and (1.46 ± 0.05cm) against Escherichia coli. Methanol and Butanol crude extract leaves of Datura showed
maximum zone of inhibition at 24hrs, 48hrs and 72hrs (3.6 ± 0cm), (3.6 ± 0.1cm) and (3.8 ± 0.05cm) against
Paenibacillus barcinonesis. Datura seed methanol crude extract crude extract produced maximum zone of inhibition
at 24hrs, 48hrs and 72hrs (1.06 ± 0.28cm), (1.5 ± 0.05cm), and (1.56 ± 0.05cm) respectively against Escherichia
coli. Datura seed methanol crude extract crude extract produced maximum zone of inhibition at 24hrs, 48hrs and
72hrs (1.33 ± 0.05cm), (1.53 ± 0.15cm), and (1.6 ± 0.2cm) respectively against Paenibacillus barcinonesis.
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Antimicrobial Pics
Datura Leaves & Datura Seed Paenibacillus barcinonesis
Neem Leaves & Bark Paenibacillus
barcinonesis
Orange Peel & Waste Paenibacillus barcinonesis
Datura Leaves & Seeds E.coli
Neem Leaves & Bark E.coli
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Orange Peel & Waste E.coli
Graphs
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IV CONCLUSION
From the above study, it concludes that the presence of constituents revealed in the solvent crude extract of leaves of
Datura, Neem, Orange could contribute for their antimicrobial activities. The various solvent extracts of the plant
showed high potential of antibacterial activities against the tested microorganisms.
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[3]Christhudas, N.I.V.S., Kuar, P.P.and Agastian,P.(2012).Antimicrobial Activity and HPLC Analysis of
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[4]Harborne, J. B. (1998). Phytochemical methods. 3rd
edition. Chapman and Hall (eds.).U.K. Pp. 74-203.
Iranbakhsh, A., Ebadi, M. and Bayat, M. (2010). The Inhibitory effects of plant methanolic extract of Datura
stramonium L. and leaf explant callus against bacteria and fungi. Global Veterinaria. 4 (2): 149-155.
[5]Johnson, M., Wesely E.G., Selvan, N. and Kavitha, M.S. (2010). In vivo and invitro anti-bacterial efficacy of
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