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Product Category: Projects

Product Code: 00000469

No of Pages: 49

No of Chapters: 5

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To investigate in vitro anti microbial activity of aerial parts of Calotropis procera and Centella asatica Linn., that have been popularly used as folk medicines. The organic solvent plant extracts are tested on the various micro-organisms including bacteria and fungi using agar well diffusion technique. The length of the inhibition zone was measured in millimeters from the edge of the well to the edge of the inhibition zone. Calotropis procera showed significant to moderate activity against (14mm) Pseudomonas marginalis and (21mm) Streptococcus mutans with 100mg/ml DMSO plant drug concentration. The results of (MICs) values are lowest at 66 and highest at 152mg/ml for Calotropis asatica. The extracts were assessed in an effort to validate the potential activity of the medicinal plant plants against microbes.










Title page                                                                                          i

Certification                                                                                      ii

Dedication                                                                                         iii

Acknowledgement                                                                             iv

Abstract                                                                                             v

Table of Content                                                                                vi-vii



1.0     Introduction                                                                            1-6



2.0     Literature Review                                                                     7

2.1     Local Names                                                                            7

2.2     Botanical Description                                                              7-8

2.3     Biology                                                                                    8

2.4     Ecology                                                                                    8

2.5     Biophysical Limits                                                                             8

2.6     Documented Species Distribution                                           8-9

2.7     Exotic                                                                                      9

2.8     Products                                                                                  9-11



3.0     Materials and Methods                                                            12

3.1     Plant Materials and Extraction                                                12-13

3.2     Determination of Antimicrobial Activity                                   13-15

3.4     Results and Discussion                                                           15-20



4.0     Result and Discussion                                                             21

4.1     Proximate Analysis                                                                  21

4.2     Determination of Moisture Content                                         21

4.3     Determination of Crude Protein                                               21-22

4.4     Determination of Ash Content                                                 22-23

4.5     Determination of Fat Content                                                  23-24

4.6     Determination of the Crude Fiber                                            24-25

4.7     Determination of Carbohydrate or Nitrogen Free Extract         25-27



5.0     Recommendation and Conclusion                                           28

5.1     Recommendation                                                                     28

5.2     Conclusion                                                                              28

References                                                                               29-30












According to a study conducted by the World Health Organization (WHO) based on publications on pharmacopoeias and medicinal plants in 91 countries, the number of medicinal plants is nearly 21,000. Nearly 6-7 thousand species of medicinal plants out of about 17-18 thousand flowering plants are known to be in use in folla and officially recognized systems of medicine in India i.e. Ayurbeda, Sidha, Unani and Homeopathy.

India has a rich heritage of knowledge on plant based, drugs both for use in preventive and curative medicine. From ancient civilization the various parts of different plants were used to eliminate pain, control suffering and counteract diseases. Plants generally produce many secondary metabolites which constitute an important source of microbicides, pesticides and many pharmaceutical drugs. It has also been widely observed and accepted that the medicinal value of plants lies in the bioactive phytocomponents present in the plants (Veeramuthu et al; 2006, Khan and Khan, 2008, Ghosh et al; 2008, Mandal et al; 2010, Abubakar et al; 2011). Much work has been done on ethnomedicinal plant in india (Nair 2005; Rout et al; 2009, Madhuri Sharma and Pandey Govind, 2009). Medicinal plants represent a rich source of antimicrobial agents (Kannan et al; 2007); Jeeshna et al; Jarrar et al; 2010, Johnson et al; 2010, Koochak et al; 2010). Because of the side effects and the resistance that pathogenic microorganisms build against antibiotics, many scientists have recently paid attention to extracts and biologically active compounds isolated from plant species used in herbal medicine (Mst Nazma Yesmin et al; 2008).

Calotropis procera linn. (Family Asclepiadeceae) is a shrub or small tree up to 2.5 m height. All parts of the plant exude while latex when cut or broken. It is known by various names like swallow wort, dead sea apple, Sodom apple or milk weed, commonly used and known as Arka or madar. Telugu name is Jilledachetta and in English Catotrope, Calotropis, Deadsea fruit, desert wick, giant milk weed, muder fibre, rubber bush, rubber tree, Sodom apple, swallow, wort. In the traditional Indian medicinal system, it has been used for pain, asthma, bronchitis and dyspepsia, leprosy, Ulcers, tumors, piles, spleen, liver and abdomen. The root bark in the form of paste with rice vinegar is applied to elephantiasis of the legs and scrotum. The plant is also known for its toxic properties that include iridocyclitis, dermatitis and acts like a poison and produces lethal effects. Compounds derived from the plant have been found to have emeto, cathartic and digitalic properties. Antimicrobial activity of C. procera was previously screened by (Kareem et al; 2008; Verahalarao Vadlapudi et al; 2004). In the leaves, mudarine is the principal active constituent as well as a bitter yellow acid, resin and 3 toxic glycosides calotropin, Uscharin and Calotoxin. The latex contains a powerful bacteriological enzyme, a very toxic glycosides calactin (the concentration of which is increased following insect or grasshopper attack as defense mechanism), Catatropin D1, Calotropin DII, Calotropin F1, Calotropin F11, and a non toxic protealytic enzyme calotropin (2 % to 3 %). This calotropin is more proteolytic than papain and bromelain coagulates milk and digest meat, gelatin and casein. The whole plant contains a b-amyrin, teraxasterol, gigcintin, giganteol, isogiganteol, b-sitosterol and a wax.

Centella asiatica (Family Mackinlayaceae) common name include Gotukola, Asiatic pennywort, Indian Pennywort, Luei Gong Gen, Takip, Kohol, Antanan, Pegagan, Pegaga Vallaarai leula kud, Baibua bok and Brahmi. In India, famous as “Brahm”, it is used for improving the mental ability as was carried out at Dr. A Laleshmipathy research center (now under CCRAS) VHS, Adyar, Chennai (Appa Rao et al; 1973). Anti bacterial activity of C. asiatica was previously screened by (Rohimi Kiran Kunta et al; 2004).

The aim of the present study is to investigate in vitro antimicrobial activity of Indian medicinal plants Calotrophs procera and Calotrophs Asatica linn against the resistant pathogens.

Fig 1: Antimicrobial activity of methanol extracts of Calotrophs procera


Fig 2: Antimicrobial activity of methanol extracts of Calotrophs asiatica

Other pathogens F. Oxysporum, L. acidophilus, S. saliuarious and S. aureus with 100mg/ml DMSO.

The results of MICs values are lowest at 66 and highest at 152 mg/ml for Calotrophs procera whereas 0.155mg/ml for Calotropis asatica. The variation of antimicrobial activity of our extracts. No variation of antimicrobial activity of our extracts might be due to the distribution of anti microbial substances which varied from fraction to fraction of the crude extract. No inhibitions were observed were observed with Calotropis procera on P. expansum and Calotrophs asatica on P. aeruginosa and S. mutans.

These extracts are harmless and nonphytotoxic, it has been proved that extracts have inhibitory effects on germination and on the viability of fungal spores as well. Both plant extracts showed moderate to good activity against A. niger as it is a saprophyte in soil causing black mould of Onion, garlic and Shallot, stem rod of Dracaena, root stalla rot of Sanseviera, and boll rot of Cotton; spoilage of Cashew kernels, dates, Figs. Vanilla pods and dried prune.

The effectiveness of the active compounds present in plant extracts causes the production of growth inhibition zones that appear as clear as surrounding the wells. However, plant extracts was unable to exhibit antibacterial activity against tested bacterial strains. These bacterial strains may have some kind of resistance mechanisms e.g enzymatic inactivation, target sits modification and decrease intracellular drug accumulation (Schwarz and Noble, 1999) or the concentration of the compound used may not be sufficient.

The adverse effect of Calotropis procera consumption are reported to cause blisters, lesions and eruptions when taken by patients for the treatment of joint pains and gastrointestinal problems. Due to its toxicity, the latex extracted from the stem has traditionally been used to make poison arroles. The latex is highly toxic to human eyes and produces sudden painless dimness of vision with photophobia (Basak et al; 2009).

Several phytochemicals are identified in different parts. Calotropis procera flowers contain terpenes, multiflorenol, and cyclisadol (Al-yahya et al; 1990). The latex contains caoutcholic calotropin, calotoxin 0.15%, Calactin 0.15%, Uscharin 0.45%, Trypsin, Voruscharin, Uzarigenin, Syriogenin and proceroside (Atef et al; 1999). Chemical constituents of calotrophs procera flowers are lupeol, Uscharin, proceroside, proceragenin (Cardenolide), Syriogenin, Taraxase-20 (30)-en-3-(4-methyl-3-periotenoate), 3-thiazoline cardenolide, gigantin, giganteol, isogiganteol, Uscharidin, Uzarigenin Voruscharin a, Calotropeol, 3-epimoretenol, a-lactuceryl acetate and alactuceryl isovalerate (Ansari and Ali, 199). Root bark of Calotropis procera contains triterpenes, A new norditerpenyl ester, name calotropterpenyl ester, and two unknown pentacyclic triterpenoids, namely Calotropursenyl acetate and Calotropfriedelenyl acetate, akundarol isovaleratae, mundarol isovalerate and quercetin, 3-rutinoside (Akhtar and Malik, 1998; Ansari et al, 2001). The principal active medicinals are asclepin and mudarin (Raghubir et al; 1999).

No inhibition was observed with controls, which proves that solvents could not act as antimicrobial agents. In almost all test, crude methanolic extracts showed better inhibition against all the tested bacterial and fungal strains, indicating that active ingredients in plant materials could be extracted into methanol.

However, the highest antibacterial activity of Calotropis procera was observed due to the presence of secondary metabolites such as alkaloids, flavanoids and steroids against S. aureus.

P. aeruginosa are wide spread in soil, water and sewage and this can be considered as an indication of their involvement in the natural process of mineralization of organic matter. It has long been a troublesome cause of secondary infections of wound, especially burns, giving rise to blue green pus. It produces meningitis, when introduced by lumber puncture and urinary tract infection when introduced by Catheters and instruments or irrigation solutions (Willey et al; 2008).

S. aureus occur harmlessly as a normal flora of the skin and mucous membrane and it is one of the commonest bacterial pathogens encountered in the community causing severe food poisoning or minor skin infections to several life threatening infections (Brooks et al; 2004).

Calotrophs asiatica methanol extract having strong inhibition activity against P. aeruginosa and S. aureus was previously reported by (Thangavel Arumugam et al; 2001)

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