Project Neem Sanjay

8th Sem

Project Work on Neem

General Introduction: Introduction:

Azadirachta indica) indica ) is one of the most suitable and valuable tree species found in Neem ( Azadirachta India. It can grow on wide range of soils upto pH 10 which makes it one of the most versatile and important trees in Indian sub-continent. Due to its multifarious uses, it has been cultivated cultivated by Indian farmers since vedic period and it has become now part of Indian culture. In India, it occurs throughout the country and can grow well in every agro-climatic zones except in high and cold regions and dam sites. In fact in India, Neem trees are often found growing scattered in the farmers fields and on the boundaries of fields without much affecting the crops. Farmers practice this system just to meet the local demand for timber, fodder, fuelwood fuelwood and also for various medicinal properties. properties. Due to its deep tap root system, it does not compete with annual crops for scarce soil moisture. Neem tree can be labeled as wonder tree for its multipurpose multipurpose uses in real sense. This has been used as a medicinal plant for long time and provides almost all the requirements of rural areas - be the timber, fuel wood, fodder, oil, fertilizers, pest repellant or the ubiquitous 'datun'.

Today, it has been recognised as the most potential tree of India due to its ever green nature (deciduous in drier areas) and ability to grow in even the most arid and nutrient deficient soils as well as for its many commercially exploitable by-products and environmentally beneficial characteristics (it has therefore been labelled as tree of the future). If plantation of this tree has to be taken up on large scale, it has to be integrated as an important component of agriculture under various agro-forestry systems. It has been estimated that India's Neembear about 3.5 million tonnes of Kernels every year. From this about 7 lakh tones of oil might be recovered. The annual production in the late 1980's was only around 1.5 lakhs tones. To increase the amount of oil harvesting, Khadi and Village Industries Commission (KVIC) has pioneered various aspects of processing the fruit and seeds of neems over the past two decades. The major difficulty as observed in most of the tree borne oil seeds including neem is that neem fruits must be harvested during the wet season. Without locally available drying facilities the fruit and seeds rapidly deteriorate and become contaminated with aflotoxin. Ideally, the fruits should be depulped without delay and the seeds have to be thoroughly dried. KVIC has popularized simple methods for depulping, drying and decorticating neem products even in the rearmost villages of the country. The sales and turnover of neem seeds in India has been estimated by various agencies. Based on random survey at major neem seeds market by independent agencies the quantity of neem seed sold during 1996 was 5.5 lakh tonnes with turnover of Rs.137 crores.

SANJAY SINGH B.Pharm

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History of Neem -:

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(Mrs. Farzana Panhwar, May 2005)

Indian scientist started neem research in 1920‟s. Neem‟s ability to repel insects was first reported in the scientific literature in 1926 and 1929. In 1962, three scientists showing that neem applied to vegetable crops would repel locusts. 1968 United Nations report called neem plantation in northern Nige ria” the greatest boon of the century” to the local inhabitants. Researchers at the U.S. Department of Agriculture have been studying since 1972. Neem‟s insect-growth-regulation insect-growth-regulation (IGR) effects were independently observed in England and Kenya in 1972. 1975 U.S. Department of Agriculture Research at Beltsville, Maryland and 1919 of its stations across the country embarked on a comprehensive program to study the post-control properties of the various plants. Bangladesh, Burkina-Faso, Chad, Myanmar, Senegal, Thailand and Mali are being assisted by Denmark and France to set-up an International International Net Work. New Guinea neem was introduced at the beginning of 1980s. Queensland and New South Wales, in 1990, 16000 tree were planted the pre 1980 neem project was for ornamental or for timber but post 1980 project for pesticide. According to Morgan (1981), there is a marked difference in the yield of Azadiractin from neem ( Azadirachtaindica,A.Juss) seeds from various sources. From seeds from Ghana he obtained 3.5 gm/Kg, and from old Indian seeds less than 0.2 g/kg. 1991, several hundred researchers in at least dozen countries were studing various aspects of neem and its products. In Nicaragua neem trees have been introduced through a cooperative known as “COPINIM” helped helped by Germany, Australia and Sweden. In 1995 they produced 200 tons of neem fruit.















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Plant profile

(THE AYURVEDIC PHARMACOPOEIA OF INDIA)

Azadirachta indica, A. Juss Syn. Biological source-: Neem consists of dried leaves of Azadirachta

Meliaazadirachta Meliaazadirachta Linn. (Fam . Meliaceae ); a moderate sized to fairly f airly large evergreen tree, attaining a height of 12-15 m with stout trunk and spreading branches, occurring throughout the country up to an elevation of 900 m.

SYNONYMS

Sanskrit: Assamese: Bengali: English: Guajarati: Hindi: Kannada: Malayalam : Marathi : Oriya : Punjabi : Tamil : Telugu : Urdu

Arista, Picumarda Mahanim Nim, Nimgach Margosa Trees Limba, Limbado, Limado, Kohumba Nim, Nimb Nimba, Bevu, Oilevevu, Kahibevu, Bevinama Veppu, Aryaveppu, Nimbam, Veppa Balantanimba, Limba, Bakayan, Nim, Kadunimb Nimba Nimba, Bakan, Nim Vemmu, Veppu, Arulundi, Veppan Vemu, Vepa Neem















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TAXONOMIC HIERARCHY OF NEEM Kingdom: Division: Class: Order: Family: Genus: Species: Other species: Scientific NameFound In:

Plantae Magnoliophyta Magnoliopsida Sapindales Meliaceae Azadirachta A. indica A.juss A.azedarac Azadirachta indica Ranthambore National Park, Bandhavgarh national Park, Mrugavani Naional Park, Bannerghata National Park, Sariska

Description-:

Character

Colour

Leaf Slightly Yellowishgreen

Bark

Flower

Fruit

Grey or dark reddish brown

White or pale yellow

Young- RipenGreen Yellow -

Odour

indistinct

-

Honey like scent

Taste

Bitter

-

Bitter

YoungBitter

RipenSweet

A large evergreen tree, 12 to 18 meter in height and 1.8 to 2.4 meter in girth with a straight bole and long spreading branches forming a broad crown as much as 20 meters across, commonly found throughout greater parts of India. Leaves Compound, alternate, rachis 15-25 cm long, 0.1 cm thick; leaflets with oblique base, opposite, exstipulate, lanceolate, acute, serrate, 7-8.5 cm long and 1.0-1.7 cm wide, slightly yellowish-green;















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Bark Grey or dark reddish brown with numerous and scattered tubercles. The bark exudes a gum known as East India gum. Leaves alternate 20 - 30 cm long, leaflets 8 19 alternate or opposite ovate glossy, bluntly serrate. Flowers : white or pale yellow, small, scented, numerous on long auxiliary panicles, have a honey like scent and attract many bees. Fruit : Fruit is a ovoid bluntly pointed, smooth drupe green when young and turns yellow with a very thin epicarp, mesocarp with scanty pulp and a hard bony endocarp, enclosing one seed. The timber is relatively heavy with a specific gravity varying from 0.56 to 0.85 (average 0.68) when freshly cut, it has a strong smell.

The flowering season of neem varies from place to place. Generally it flowers from January to May and the ripening time of fruits is from May to August. The fruit pulp is edible.

Geographical Geographic al distribution It has been widely cultivated in India and African countries. Native of Burma but grown all over India. In India, it occurs throughout the larger parts of the country in the states of Uttar Pradesh, Bihar, West Bengal, Orissa, Delhi, Maharashtra, Gujarat, Andhra Pradesh, And Tamil Nadu.















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CULTIVATION AND COLLECTION

([email protected])

Neem tree can grow under variousconditions of climate and soil types.The ripe seeds should be grown soon after collection. It can be sown directly or it can also be transplanted. Neem can also be propagated in a variety of other ways.

Propagation Techniques of Neem The tree can be easily propagated both sexually and vegetatively. It can be planted using seeds, seedlings, saplings, root, suckers, or through tissue culture. However, it is normally grown from seed. Neem seeds are not viable for a very long time. After about two weeks they will no longer germinate. The seeds that are removed from the fruit should be put in water. Those seeds which sink are good ones and should be used. Some studies have shown that they can be used even after many years but only under specific condition and germination capacity is under 50%. Neem cuttings have to be planted as soon as they are cut for the neem to survive. The part that is cut is covered with straw and should be kept wet if it is to be planted after some time.

Preparation of nursery and Sowing The soil should be dug 30 cm deep. Raised beds of 10m length, 1 m breadth and 15 cm height should be prepared. Farmyard manure(FYM), sand and local soil should be mixed in the ratio 1:1:3. This should be put on top of the soil for a height of 2.5 to 5 cm. Seeds are normally sown in the nursery in the months of July and August. The seeds should be sown at a distance of 15 to 20 cm and a depth of 1 to 1.5 cm, and then watered. Hence they can be sown as soon as they are collected. They germinate in a week's time.After 5 to 6 weeks the

















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bags at the rate of 2 per bag. The healthy seedling is maintained and the other one removed. If the seeding are raised in the polythene bag they can be transported for long distances when

required and then transplanted. The polythene bags that are used should be 150-200 gauge in weight. In the nursery a thatch should be provided for seedlings. Transplantation The place where neem seedlings are to be transplanted should be properly ploughed. The seedlings are ready for transplantation when they are six months old (15 to 22.5 cm height). If the seedlings which are kept for a long period the tap roots become very long and it is difficult to take out without breaking of roots. Seedlings which are healthy and not afflicted with any diseases should be chosen. Pits of dimension 30x30x30 cm should be dug at intervals of 3x3 m. The neem seedling should be transplanted during the period of the South East Monsoon. This increase their survival rate. Initially, Initially, they should be irrigated once in two or three days interval. After they grow well they can be wanted once in seven to ten days.

Harvesting In most parts of India today, leaves are collected collected by simply cutting or picking picking method and separated from leaflet then dried in shade avoiding any contamination .leaves are generally collected when they are neither yellowish nor green. Seeds that fall on the ground are simply collected. Neem fruit fall coincides with the monsoon. At moisture content above 14%, neem fruits carry the fungus Aspergillus flavus. These funguses under many conditions produce aflatoxins. Aflatoxins are extremely potent among carcinogens known and these could also contaminate seeds inside the fruits. They are extracted and concentrated along with pesticidal ingredients. Seed viability is also lowered. To avoid all this actual harvesting of seeds is recommended rather than a simple collection of seeds. While harvesting neem seeds it must be ensured that the fruit colour is neither greenish yellow nor brownish yellow but plain absolute yellow. Greenish yellow fruit are not fully mature and are low in Azadirachtin content. After indentifying trees which have yellow fruit a tarpaulin cloth or plastic paper is spread under the tree. A stick is used to beat the branches the same way as one would to collect Tamarind.

















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Traditional Uses:

(P. Sudhir et al )

Various parts of the neem tree have been used as traditional ayurvedic medicine in India from time immemorial. The medicinal utilities have been described, especially for leaf, fruit and bark. 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. Its use for the treatment of rheumatism, chronic syphilitic sores and indolent ulcer has also been evident. 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 pthysis. Some of the medicinal attributes of various parts of neem as mentioned in ayurveda have been summarized in Table 2. However, apart from these uses, there are several reports on the

















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The aqueous extract of neem bark possesses anticomplement activity, acting both on the alternative as well as the classical pathway of complement activation in human serum. Recently, an aqueous extract of stem bark has been shown to enhance the immune response of Balb-c mice to sheep red blood cells in vivo. The aqueous extract of leaf also possesses potent immunostimulant activity as evidenced by both humoral and cellmediated responses. Leaf extract at 100 mg/kg after three weeks of oral administration causes higher IgM and IgG levels along with increased titer of antiovalbumin antibody. Neem oil has been shown to possess immunostimulant activity by selectively activating the cellmediated immune mechanisms to elicit an enhanced response to subsequent mitogenic or antigenic challenge. Hypoglycemic activity: Aqueous extract of neem leaves significantly decreases blood sugar level and prevents adrenaline as well as glucose-induced hyperglycemia. The aqueous leaf extract when orally fed, also produces hypoglycemia in normal rats and decreased blood glucose levels in experimentally-induced diabetes in rats. Aqueous leaf extract also reduces hyperglycemia in streptozotocin diabetes and the effect is possibly due to presence of a flavonoid, quercetin. A significant hypoglycemic effect was also observed by feeding neem oil to fasting rabbits. Recently, hypoglycemic effect was observed with leaf extract and seed oil, in normal as well as alloxan-induced diabetic rabbits. The possible mechanisms underlying the hypoglycemic activity of the aqueous leaf extract have also been discussed. Antiulcer effect: Neem leaf aqueous extract produces antiulcer effect in rats exposed to restraint cold stressor ethanol orally by preventing mucus depletion and mast cell degranulation. An aqueous extract of neem bark has been shown from our laboratory to possess highly potent antacid secretory and antiulcer activity and the bioactive compound has been attributed to a glycoside. Antifertility effect: Neem oil proved spermicidal against rhesus monkey and human spermatozoa in vitro. In vivo studies showed that intravaginal application of neem oil prior to coitus can prevent pregnancy. Antifertility effect of neem oil has also been studied and suggested to be a novel method of contraception. Oral administration of aqueous extract of neem leaf a antifertility effect in mice. Purified neem seed extract (Praneem) has also been demonstrated to abrogate

















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sensitive strains of malarial parasite. Recently, neem seed extract and its purified fractions have been shown to inhibit growth and development of asexual and sexual stages of drug sensitive and resistant strains of the human malarial parasite P. falciparum. Antifungal activity: Extracts of neem leaf, neem oil and seed kernels are effective against certain human fungi, including Trichophyton, Epidermophyton, Microsporum, Trichosporon, Geotricum and Candida. High antimycotic activity with extracts of different parts of neem has already been reported. Antibacterial activity: Oil from the leaves, seeds and bark possesses a wide spectrum of antibacterial action against Gram-negative and Gram-positive microorganisms, including M . tuberculosis and streptomycin streptomycin resistant strains. In vitro, it inhibits Vibrio cholerae, Klebsiella pneumoniae, M. tuberculosis and M . pyogenes. Antimicrobial effects of neem extract have been demonstrated against Streptococcus mutans and S. faecalis. NIM-76, a new vaginal contraceptive from neem oil showed inhibitory effect on the growth of various pathogens, including bacteria, fungi and virus. Recently, the antibacterial activity of neem seed oil was assessed in vitro against 14 strains of pathogenic pathogenic bacteria. Antiviral activity: Aqueous leaf extract offers antiviral activity against Vaccinia virus, Chikungemya and measles virus in vitro. The antiviral and virucidal effects of the methanol extract of neem leaves (NCL-11) have recently been demonstrated against group-B Coxsackie viruses. NCL11 inhibits plaque formation in different antigenic types of Coxsackie virus B at a concentration of 1 mg/ml at 96 h in vitro. Further studies indicated that NCL-11 is most effective in Coxsackie virus B-4 as a virusidal agent, in addition to its interference at the early events of its replication. Anticarcinogenic activity: Neem leaf aqueous extract effectively suppresses oral squamous cell carcinoma induced by 7,12-dimethylbenz[a]anthracene (DMBA), as revealed by reduced incidence of neoplasm. Neem may exert its chemopreventive effect in the oral mucosa by modulation of glutathione and its metabolizing enzymes. That neem leaf extract exerts its protective effect

















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Antioxidant activity: The antioxidant activity of neem seed extract has been demonstrated in vivo during horse grain germination, which is associated with low levels of lipooxygenase activity and lipid peroxides. An antioxidant principle has also been isolated, which is a potent inhibitor of plant lipooxygenases. Table 2. Some medicinal uses of neem as mentioned in ayurveda Part Leaf

Medicinal use Leprosy, eye problem,epistaxis, problem,epistaxis, intestinal worms, anorexia, biliousness, skin ulcers. Bark Analgesic, alternative and curative of fever. Flower Bile suppression, suppressi on, elimination of intestinal worms and phlegm. Fruit Relieves piles, intestinal worms, urinary disorder, epistaxis, phlegm, eye problem, diabetes, wounds and leprosy. Twig Relieves cough, asthma, piles, phantom, tumour, intestinal worms, spermatorrhoea, spermatorrhoea, Obstinate urinary disorder, diabetes. Gum Effective against skin diseases like ringworms, scabies, wounds and ulcers. Seed pulp Leprosy and intestinal intestina l worms. Oil Leprosy and intestinal worms. Root, bark, leaf, Blood morbidity, biliary afflictions, afflictions , Flower itching, skin ulcer, burning sensation and fruit together leprosy. __________________________________________________________________________ Effect on central nervous system: Varying degrees of central nervous system (CNS) depressant activity in mice was observed with the leaf extract. Fractions of acetone extract of leaf showed significant CNS depressant activity. activity. Leaf extract up to a dose of 200 mg/kg body weight produces produces significant anxiolytic activity in rats. The crude ethanol extract of stem bark and root bark showed hypotensive, spasmolytic and diuretic activities.

Reported Pharmacological Activities:-



















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The acidic extract from leaves and neutral extracts from seeds inhibited Ehrlich ascites carcinoma cell line growth and had anticancer activity. IC50 values of acidic extracts from leaves and neutral extracts from seeds were 669.43 and 724.63 μg/ml, respectively.

2. Anti-Hepatotoxicity

(Kale et al)

Methods: Hepatotoxicity was induced in rats by combination of isoniazid, rifampicin and pyrazinamide given orally as suspension for 30 days. Treatment groups received AI aqueous leaf extract along with ant tubercular drugs. In the second phase of study the effect of aqueous leaf extract on established hepatotoxicity was studied by giving the extract for 20 days after withdrawal of antitubercular tubercular drugs. Liver damage was assessed by biochemical and histological parameters. Conclusion: Aqueous leaf extract(Neem) significantly prevents and reverses the hepatotoxic damage induced by antitubercular drugs in rats. 3.

Anti-fertility activity-:

( Indian J. med. Res. 1983)

Oral administration of 20ml of neem oil from day 1 to 10 of pregnancy to female rats, th showed on 16 day growth of fetuses retarded in 50% animals. In another group of animals given the same treatment orally but allowed to complete the term, 80% animals

















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Extract of leaves did not show oestrogenic activity in rats, as evidenced by insignificant change in weight of uterus and adrenal, absence of open vagina and epithelial cells in vaginal smear and incomplete development of uterus. 6. Insecticidal activity

( Heterocycles 1986), (J.Chem. soc. Perkin 1 1987)

An acidic fraction from leaves showed insect growth regulating activity in mosquitoes. Plant extract showed insect growth regulating activity against pulse beetle, callasobruchus analis

. 7. Anti-bacterial activity

(Phytochemistry 1988)

Purified bark extract showed significant anti-bacterial activity. 8. Anti-hyperglycemic

( Phytother.res. 1989)

Aqueous extract of leaves lowered hyperglycemia in streptozotocine induced diabetes

















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Nimbidol:Provides anti-tubercular, anti-protozoan and antipyretic properties Sodium nimbinate:Provides diuretic, spermicidal and anti-arthritic properties

Providesvasodilator, anti-malaria and antifungal antifungal properties Gedunin:Providesvasodilator, Salannin:Provides repellant properties Quercetin:Provides properties

anti-protozoal,

antioxidant,

anti-inflammatory

STRUCTURE OF PHYTOCHEMICAL CONSTITUENTS-:

and

antibacterial

















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PLAN OF WORK1. PHARMACOGN PHARMACOGNOSTICAL OSTICAL EVALUATION I.

Macroscopic evaluation

II.

Microscopic evaluation 

Leaf section, powder microscopy

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Leaf stomata

2. PHYSICOCHEMICAL EVALUATION/STUDY

















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1. PHARMACOGNOSTICAL EVALUATION (WHO GUIDELINE 1998)

I.

MACROSCOPIC EVALUATION

Character

Colour

Leaf Slightly Yellowishgreen

Bark

Flower

Fruit

Grey or dark reddish brown

White or pale yellow

Young- RipenGreen Yellow -

Odour

indistinct

-

Honey like scent

Taste

Bitter

-

Bitter

YoungBitter

RipenSweet

















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II.

MICROSCOPIC EVALUATION 

Leaf section cutting

Equipment The following are required: - a microscope equipped with lenses providing a wide range of magnification and a substage condenser, a graduated mechanical stage, objectives with a magnification of 4x, 10x, and 40x, and colour filters of ground glass, blue-green; high eyepoint eyepieces are preferred for wearers of spectacles;

a lamp, either separate or incorporated into the microscope; - a stage micrometer and an ocular micrometer to be inserted into a 6x eyepiece and placed on the diaphragm or, preferably, a micrometer eyepiece; - a set of drawing attachments for the microscope;



















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III. LEAF STOMATA In the mature leaf, four significantly different types of stoma are distinguished by their form and the arrangement of the surrounding cells, especially the subsidiary cells (Fig. 2):

















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Types of leaf stoma

















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2. PHYSICOCHEMICAL EVALUATION I.

ASH VALUE

The ash remaining following ignition of medicinal plant materials is determined by three

















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Subtract the weight of this residue in mg from the weight of total ash. Calculate the content of water-soluble ash in mg per g of air-dried material.

II.

EXTRACTION METHOD-:

This method determines the amount of active constituents extracted with solvents from a given amount of medicinal plant material. It is employed for materials for which as yet no suitable chemical or biological assay exists. Recommended procedures

















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followed by Buchner funnel and the filtrate was collected in a round bottom flask. The process was repeated using marc for for another two times times and all the three filtrates filtrates were collected and combined. The solvent was removed and concentrated in-vacuo in a rotary evaporator. The dried extracts were placed in a desiccator and used for further studies. The colour, consistency and the % yields of o f the extracts were recorded.



















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Project Neem Sanjay

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