eChapters

A family of about 244 genera and 4,225 species, Malvaceae are distributed widely in tropical and temperate regions. About 22 genera and 125 species of Malvaceae have so far been reported from India. Some of the larger
genera, along with their common names and/or number of reported species in parenthesis, include Hibiscus (rose mallow, 300), Sida (200), Abutilon (Indian mallow, 100), Alcea (60), Malva (mallow, 40), Lavatera (25), Gossypium (cotton, 20) and Althaea (12). The family is recognized by Hibiscus rosasinensis (Rose of China) because of its beautiful large flowers and hundreds of its known cultivated varieties.
Hibiscus commonly called as ‘Chinese hibiscus’ or ‘Rose of China’ or ‘Hawaiian hibiscus’ or ‘Blacking plant’ or ‘Shoe flower’ is a flowering plant of hot and warm climate, and hence, known as ‘Queen of Tropics’. Florida is the
prominent hibiscus growing centre of the world. The commercial species have wide variety of uses including fibers, food, medicinal and ornamental, either as a potted plant, fence or hedge plant or as a landscape shrub. Ethno medicinal value of hibiscus is documented

A family of about 150 genera and 3,800-5650 species, distributed mainly in sub-tropical and tropical regions of the world. Family is particularly abundant in Australia where some Eucalyptus trees are world’s tallest angiosperms. Myrtaceae is represented in India by about 15 genera and over 170 species. Beautiful pinkish flowers of Myrtus communis are considered highly sacred among Greeks, Egyptians, Jews and Persians, and used in religious rites and ceremonies. Some of the larger genera of Myrtaceae along with their number of reported species include Eugenia (1000), Eucalyptus (500), Syzygium (500), Myrcia (500), Psidium (140), Myrtus (100) and Melaleuca (100).
Eucalyptus spp. produce important components of medicinal and pharmaceutical applications. Economic parts are leaves and terminal buds-along with trunk. Lemon scented/ Tasmanian (E. globulus) is high in cineol content. Deodorant is manufactured besides utilization of oil for asthma, bronchitis, perfumery, etc. Phellandrene rich type, E. citridora (most widely grown in hill stations of southern parts of India). The source of citronellal and menthol from leaves and terminal buds is famous for perfumery industries.

This family of sub-class polypetalae is represented by only single genera and three species. Members are cosmopolitan in distribution, except in very cold regions, and are favorite plants for garden pools. Nelumbo lotus are the ‘sacred lotus’ plants of the family. Indian lotus, found commonly in shallow lakes and pods, is Nelumbo nucifera. Nelumbo, genus of aquatic angiosperms belong to monogeneric family Nelumbonaceae is represented by two elemental species N. nucifera and N. lutea. The 1st fossil record available with Pleistocene epoch of Kashmir.The NBRI, Lucknow acts as a biodiversity conservation centre.
Among the few bulbous ornamentals which survive only in water, one of them is lotus.N.aureavallis, known from Eocene fossils from North Dakota, USA is an extinct species from USA. Nearly all parts are edible. Leaves are used for wrapping around other food before it is cooked. Stalks are sliced & added to salad. Flowers are cultivated for ornamental value, religious offerings, environment beautification, water purification, besides for food, Chinese traditional medicine & yield valuable perfume. Flowers recommended as cardiac tonic and used in curing diseases of liver, skin. Its dried and powdered rhizomes are used to cure piles. Due to its astringent qualities, rhizome is known for its hypoglycemic, anti-pyretic, anti-diarrheal, anti-bacterial and anti-inflammatory activity. It possesses anti-fungal activity against Aspergillus, Trichoderma, and Penicilium. Many traditional ancient medical texts report its use for skin conditions, notably ring worms & leprosy, sexually transmitted diseases such as gonorrhea and syphilis. Milky latex found in stems, leaves and flowers is used to fight bacterial infections. Lotus has been found to have 77% anti-oxidant activity. Swollen rhizome of vegetable lotus is used in culinary preparations. The roasted seeds are used as a substitute to coffee. Seeds are ground to obtain almond-flavoured flour. Seed pods (fresh or dried) are used in dry flower arrangement. Bhasinda is an underground stem i.e. rhizomes are edible as raw or cooked in northern parts of India.

A family of over 33 genera and 391 species, Nyctaginaceae are distributed mainly in tropics, but also in South America particularly Brazil, Asia, Ecuador, Peru, Columbia, Australia, Africa and New Zealand are the chief centers of distribution. Presently, bougainvilleas are popular ornamental plants in warm climates including India, Taiwan, Vietnam, Malaysia, Australia, Mediterranean region, the Caribbean, Mexico, South Africa, and the United States in Arizona, California, Florida, Hawaii, and southern Texas. Some of the largely represented genera along with the approximate number of their world over reported species are Bougainvillea (300), Mirabilis (60) and Pisonia (50). The C3 physiology recorded directly in Bougainvillea, Mirabilis.
Bougainvillea is a versatile and hardy climbing shrub commonly grown in Indian gardens. It is a short day plant. C3 and C4 photosynthesis is reported in bougainvillea cv. Mary Palmer. It is a fast growing garden plant admired for its brightly coloured bracts, profuse flowering round the year, wide adaptability to soil and climatic conditions, and easy growing. It is, grown as hedge, pergola, standard, pot plant, and bonsai for display. In city designs, newer roads, and flyovers are introduced with bougainvilleas as dividers. Bougainvilleas also have been utilized as a kind of folk medicine developed into dietary supplements as insulin substitute and for pollution tolerance. The insulin like compound, pinitol, was first isolated from leaves of B. spectabilis.

This family of sub-class polypetalae is represented by only 5 genera and over 70 species. Members are cosmopolitan in distribution, except in very cold regions, and are favorite plants for garden pools. Plants are mostly ornamental, few are medicinal or edible. Flowers of water lily are variously coloured and open just above the water surface in ponds while that of crow lily (Nuphar) are golden yellow and extend up and out of the water in the pond. Nymphaea nouchali is the Indian red water-lily. Victoria regia is a native of South America. Genus Nymphaea attained wide spread fame as ornamentals in Europe and North America in the early 19th century. Word ‘Nympha’ in Greek and in Roman mythology means ‘Nature-goddess’. Nymphacine is an alkaloid present in all parts except seeds and contain 47% starch and fatty oils. Flowers are used as cardiac tonic to relieve pains and cures diseases of liver and skin and rhizomes are used in tannin industry, seeds for culinary purposes and leaves as dining plates. Dried rhizome powder is used to cure piles.
Among the few bulbous ornamentals which survive only in water, one of them is water lily. Water lilies constitute an important element in tropical aquatic ornamental horticulture because of their strikingly beautiful flowers with wide spectrum of colours. Its flowers are very formal in shape and rather more attractive than lotus. Many are fragrant and there is great variation even in size of the flowers and leaves. Unripe fruits of Nymphaea rubra are used as vegetables and flowering stalks as vegetables and salad. Rhizomes are used in tannin industry and seeds for culinary purposes. Leaves are used as dining plates. Nymphacine alkaloid is available in all parts of the plant except the seeds which are free from this alkaloid and contain 47 per cent starch and fatty oils. Rhizomes of N. stellata are edible and also used to relieve cardiac pains. Rhizomes of N. nouchali are used to cure dysentery and diarrhoea. The vegetable ‘kamal kakri’ is actually the rhizome of N. nouchali.

A family of approximately 29 genera and 600 species, Oleaceae are cosmopolitan but occur in great diversity in temperature and tropical Asia. Some of the largely represented genera of olive family and their approximately
reported species are Jasminum (300), Chionanthus (125), Linociera (80), Fraxinus (70), Ligustrum (40), Syringa (30) and Olea (20).
Genus Jasminum contains 90 species to be true in existence and use, range from landscape to hair adornments, and perfumes. It is native to Indo-Malayan region. Morphological variants from different species have been developed in different regions in India. Jasmines are one of the most important ornamental flowering plants, widely cultivated for their attractive and fragrant flowers. Jasmine has multiple uses like in the making of bouquets, garlands, decorating women’s hair as veni, beautifying pandals and marriage halls and the like, besides preparation of perfumes and cosmetics. Jasminum grandiflorum is highly priced for high grade perfumes throughout the world. Jasmine oil has a significant place as an ingredient in cosmetics, confectionary, ointments, disinfectants and detergents and in pharmaceutical beverages. The word ‘jasmine’ has been derived from a Persian name ‘Yasmun’ meaning fragrance.

Orchidaceae is one of the largest families of the flowering plants, represented by about 1,000 genera and 20,000 species which are cosmopolitan but primarily distributed in tropical areas. They are found in all parts of the world, even in the frozen areas of Alaska, the snow-covered areas of the Himalayas and sandy deserts of Australia and Africa. New Guinea has more orchids, native to the island, than any other part of the world. In Indian flora, this is the second largest family having 7% of the world’s orchid genetic diversity contributing 1,330 species in 186 genera distributed mainly in Eastern Himalayas, Western Ghats and Khasi hills. Indian orchid wealth serves as a foundation of world’s orchid industry. The North-Eastern hilly regions of India, Himalayas being the cradle, comprises eight states- Arunachal Pradesh, Assam, Manipur, Meghalaya, Mizoram, Nagaland, Sikkim and Tripura- is the abode of unique orchids. Many are now endangered due to man’s invasion and “burn and grow” type of shifting cultivation. Arunachal Pradesh alone accounts for 601 species, almost 52% of the total species known in India. Arunachal’s rich and colourful orchids find a place of pride. Arunachal Pradesh is called the ‘Orchid Paradise of India’. The entire orchid family with about 601 species in Arunachal Pradesh is classified into six sub-families, 17 tribes, 24 sub-tribes and 111 genera. Vanda is found as epiphyte on mango and other trees in the plains. Aerides is found in southern and northern parts of India, Burma, Malaya and Indonesia, Vanda spp. is common in India, Sri Lanka, South East Asia and Indonesia. Calanthe spp. have a wide range from China, Japan, Vietnam to Australia, Southern parts of India and South Africa. The striking Cattleya spp. are found only in South America. Coelogyne, a genus of considerable horticultural value, is mostly represented in Nepal and Sikkim but several species are found in Bhutan and upper Myanmar, China, South-East Asia, the Philippines and Fiji. About 20% of recorded Indian taxa are endemic. Dendrobium peguanum is a rare species, Rhizanthella gardneri is endemic to Western Australia which grows underground and Phaius tankervilliae (Nun’s orchid of Assam; 2n= 50) is also an endangered orchid. India has two conservation and germplasm centers at Shillong in Meghalaya and Yercaud in Tamil Nadu. Asia’s largest orchidarium is situated at Tipi, Arunachal Pradesh while orchids sanctuary is situated at Sikkim.

A family of 183 genera and over 2,600 species distributed widely in tropical and sub-tropical regions of the world and a few also in warm, temperate regions. About 28 genera and over 95 species have been reported from India. Chief genera along with the number of their total reported species and the common names of them are Calamus (300, cane palm), Chamaedorea (130), Areca (54, betel nut plam), Coccothrinax (50, Biscayne palm), Copernicia (30, wax palm), Raphia (30, wine palm), Sabal (25, cabbage palm), Phoenix (17, date palm), Roystonea (17, Royal palm), Metroxylon (15, sago palm), Phytelephas (15, ivory-nut palm), Caryota (12, fish-tail palm), Elaeis (2, oil
palm) and Cocos (1, coconut palm). Selected Indian Genera Araca, Borassus, Calamus, Cocos, Nypa, Phoenix.
Economic Importance
Palm family, supposed to be second in importance to Poaceae, is used for food, oils, shelter, coconuts, copra, dates, clothing, etc. As many as 801 uses of only Borassus flabellifer have been mentioned in one ancient Tamil song. Cocos nucifera (coconut Fig. 45.2) is another such plant of this family whose almost every part of entire plant body is utilized in some ways or other, i.e.

Primrose family is represented by about 58 genera and 2,600 species which are cosmopolitan but chiefly distributed in northern hemisphere. Some of the
larger genera with their approximately reported species include Primula (500), Lysimachia (200), Dodecatheon (50), Dionysia (41), Anagallis (30), Cyclamen (15), Salomus and Soldanella (11).
Selected Indian Genera
Primula, Anagallis, Androsace, Cyclamen, Primula.
Economic Importance
The family is of no specific economic importance except that of some well known decorative plants it contains. Some species of Primula may cause contact dermatitis. Anagallis arvensis is used for curing snakebite, leprosy, gout and even hydrophobia. Cyclamen purpurascence contains glucoside cyclamen which is poisonous. Some of the commonly cultivated ornamental plants of Primulaceae are the species of Primula (primrose), Androsace (rock jasmine), Cyclamen (Fig. 46.1), Dodecatheon (shooting star), Douglasia, Lysimachia (Loosestrile) and Omphalogramma.

This family is represented by about 62 genera and 2,252 species, chiefly distributed in the temperate regions of the Northern Hemisphere. Eastern Asia and eastern North America are the regions of their most common occurrence. The common genera, of which over 150 or more species have so far been reported, include Ranunculus (400), Aconitum (300), Delphinium (250), Clematis (250), Anemone (150) and Thalictrum (150). Some species occur in Arctic and Alpine zones. However, they are rare in tropics. Twenty genera and over 165 species of this family have so far been reported from India. Plants of this family flower mainly during winter in our country. Selected Indian Genera and Species
• Ornamentals: Many Ranunculaceae have showy flowers and are grown as ornamentals in gardens. Some of such ornamentals are Aconitum napellus (aconite), Anemone parviflora (wind flower), Aquilegia coerulea (columbine), Caltha palustris (Marsh marigold), Clematis virginiana (traveller’s joy), Delphinium ajacis (larkspur), Helleborus (Christmas rose), Nigella damascena (love-in-a-mist), Paeonia emodi (Peony), Ranunculus aquatilis, R. seleratus (cedery peaved crow foot).

A family of approximately 91 genera and 4,828 species which are cosmopolitan in distribution but particularly abundant in Eastern Asia, North America and Europe. About 25 genera and 230 species of Rosaceae have so far been reported from India. Some of the largely represented genera along with their common names and approximately reported species include Potentilla (Cinquefoil, 500), Prunus (peach, plum, almond, cherry, apricot, 430), Rosa (Rose, 250), Rubus (Raspberry, blackberry, 250), Alchemilla (250), Crataegus (Hawthorn, 200), Sorbus (Chokeberry, 250), Acaena (100) and Spirea (100). Named after the universally famous flowering plant, rose, the Rosaceae show great diversity in types of fruit (Fig. 48.1).
Roses, universally acclaimed as the ‘queen of flowers’, have played major role in human culture and artistry from ancient times. Among the ornamentals, roses occupy very important position both as garden plants and cut flowers for their beauty and fragrance. This unique position is attributed to great amount of variation in flowers and plant characteristics as well as their wide adaptability to varied agro-ecological conditions. Damask rose (R. damascena) and Edouard rose (R. bourboniana) are cultivated for extraction of essence, rose attar or rose oil, otto and other products, gulkand, gulab jal and pankhuri from petals. Rose oil has a significant place as an ingredient in pharmaceutical preparations viz., ointments, lotions, perfumes, creams and soaps and in food products including beverages, baked foods, puddings and frozen dairy desserts. Characteristics of damask rose oil as anti-HIV, anti-bacterial and anti-oxidant have been established.

A family of about 611 genera and 13,150 species, Rubiaceae are chiefly distributed in tropics, but some are distributed in temperate or even arctic regions. About 75 genera and 275 species have been reported from India, chiefly in tropical and sub-tropical eastern Himalayas. Some of the largely represented genera with their approximately reported species (Hickey and King, 1988) include Psychotria (700), Galium (400), Ixora (400), Gardenia (250), Mussaenda (200), Asperula (200), Cephaelis (180), Rondeletia (120), Coprosoma (90), Guettarda (80), Rubia (60), Coffea (40) and Cinchona (40). Two most important plants of the family are Coffea arabica (coffee-yielding plant) and species of Cinchona (quinine-yielding plant).
Economic Importance of Selected Genera and Species
Some of the common ornamental plants grown in gardens and roadsides include Anthocephalus cadamba (Kadam), Cephalanthus occidentalis, Galium verum (Ladies Bedstraw), Gardenia jasminoides (Jasmine), Hamelia patens, Hamiltonia suaveolens (Padera), Ixora arborea, I. coccinia, Morinda tinctoria, M. frondosa, M. luteola, Pentas lanceolatea and Rondeletia.

Represented by about 65 genera and 1,730 species, Scrophulariaceae are cosmopolitan in their distribution. About 60 genera and 360 species have been reported from India, chiefly from the Himalayas. Some of the genera along with their approximately reported species from world over are Pedicularis (500), Calceolaria (300-400), Scrophularia (300), Verbascum (300), Veronica (300), Penstemon (280, Linaria (150), Castilleja (150), Selago (150), Sutera (130), Mimulus (100), Antirrhinum (40) and Digitalis (20). The common names of the family (snapdragons, foxglove or figwort family) are because of its genera Antirrhinum (snapdragons), Digitalis (foxglove) and Scrophularia (figwort).
Selected Indian Genera
Antirrhinum, Digitalis, Linaria, Lindenbergia, Mazus, Verbascum, Veronica.
Economic Importance
1. Ornamental plants: Include Angelonia angustifolia, Antirrhinum majus (snapdragon), Calceolaria scabiosaefolia (slipperwort), Castilleja (Indian paintbrush), Chelone glabra (turtlehead), Collinsia grandiflora, Digitalis purpurea (foxglove), Linaria vulgaris (toadflax), Mimulus luteus (monkey flower), Nemesia versicolor (Nemesia), Paulownia tomentosa, Pedicularis canadensis (housewort), Russelia juncea (coral blow), Torenia asiatica and Veronica bungifolia (speedwell).

A family of about 90 genera and 3,000 species distributed in both tropical and temperate regions. Central and South America are the chief centres of distribution where over 40 genera are found. About 15 genera and over 90
species have been reported from India. Some of the larger genera along with their approximately world over reported species (Hickey and King, 1988) are Solanum (1700), Cestrum (150), Physalis (100), Lycium (80-90), Nicotiana (66), Capsicum (50), Petunia (40), Hyoscyamus (20) and Datura (10). Solanum tuberosum (potato) and Nicotiana tabacum (tobacco) are the two most utilized plants of Solanaceae.
Petunias, popular bedding plants, with variety of forms, sizes and shapes are employed for landscape planning, garden designs and container gardening. The numerous phenotypes viz., Grandifloras, Multifloras, Millifloras, Waves, Hedgifloras, etc. are complex hybrids. The outstanding marketable series of P. x hybrida have been developed for upright, spreading and semi-trailing to trailing habits.
Selected Indian Genera
Atropa, Capsicum, Cestrum, Datura, Nicotiana, Petunia, Physalis, Solanum, Withania.

A family of about 86 genera and 3,000 species distributed chiefly in tropics and sub-tropics. About 22 genera and over 30 species have been reported from India. Some of the genera along with their number of approximately reported species and common names of them are Clerodendron (400, glory-flower), Verbena (250, vervain), Vitex (250, chaste tree), Lippia (220, Frogfruit), Lantana (150), Callicarpa (140, beauty-berry), Stachytarpheta (100) and Tectona (3, teak).
Verbena is popular ornamental plant suitable for flower gardens, hanging baskets, and balcony boxes. They are excellent both as ground cover for rockeries, and as bedding types. These plants abundantly flower in the seedin year and, thus, are grown as annual plants.
Selected Indian Genera
Clerodendron, Duranta, Lantana, Tectona, Verbena, Vitex.
Economic Importance
1. Timber yielding plants: Teak, one of the most famous timber-yielding plants of the world, belongs to Verbenaceae. It is used for high quality furnitures, heavy constructions, ship building, musical instruments and several other similar purposes. Some other timber-yielding plants are Gmelina arborea, Premna benghalensis and Vitex altissima.

A family of about 23 genera and 800 species, Violaceae are distributed in both tropical and temperate regions. Only three genera (Hybanthus, Vahila and Viola) have been reported from India. Famous pansies, violas and violets belong to this family. Some of the larger genera with their approximate number of species in parenthesis include Viola (500), Rinorea (340) and Hybanthus (150). Majority of Indian Violaceae are found in Himalayas and hills of
southern and western India.
Garden pansy is an annual or perennial herb planted in flower beds worldwide owing to its pretty shape and contrasting colour markings, looking like a butterfly. It is called as ‘King of annuals’. In horticultural industry, there are variety of forms, sizes, and shapes of pansies which are employed for landscape planting, garden design, and container gardening. With the advent of F1 hybrids, pansies are known as high quality bedding plants. Dried flowers of Viola odorata Vern. are used against cough, influenza, and lung troubles while oil obtained from flowers is used in perfumes and flavourings.
Selected Indian Genera and Species
• Except for garden favourites, such as pansies, violas and violets, the family is of little economic importance. Over 100 species of Viola, are being grown as ornamental plants.

A family of 56 genera and over 1,300 species, Zingiberaceae are distributed in tropical regions, chiefly in Indomalaysia. About 17 genera and over 115 species have been reported from India, chiefly in Western Ghats and Eastern Himalayas. Some largely represented genera along with number of their world over reported species are Alpinia (250), Amomum (150), Costus (150), Zingiber (90), Kaempferia (70), Curcuma (60), Hedychium (50), Globba (50) and Elettaria (7).
Selected Indian Genera
Alpinia, Amomum, Costus, Curcuma, Elettaria, Hedychium, Zingiber.
Economic Importance
1. Spices and condiments: Seeds of Amomum cardamom (Badi elayachi) and Elettaria cardamomum (true cardamom), roots of Alpinia officinarum (gangal root) and rhizomes of A. gangala (Siamese ginger), Curcuma longa (turmeric) and Zingiber officinalis (ginger) are used as spices, condiments and flavouring agents. Ginger oil, obtained from rhizomes of Z. officinalis, is used in perfumery and for medicinal purposes. Turmeric is also used for medicinal purposes.

Amaranthus is an important leafy vegetable and is grown all over the Indian subcontinent. Amaranthus tricolor Linn and A. blittum Linn. are the common varieties cultivated. These herbaceous annuals are a rich source of proteins, vitamins A and C, and iron. The foliage is a good source of calcium, iron, and vitamins A, B, and C. In contrast, grains are good sources of important amino acids, particularly lysine, and are rich in dietary fibre, calcium, and minerals such as iron, magnesium, phosphorus, copper, and manganese. It enhances the body’s immune system and antioxidant status while lowering blood pressure and cholesterol. Fresh leaves and tender stems are used as vegetables. Production of amaranthus is affected by the incidence of pests. Cultivated amaranth is currently infested by a total of 92 different insect pests. (Rajeshkanna et al., 2017) among which the leaf webbers and amaranthus stem weevil are the most important pests.
Biology: Black moth with dark brown wings bearing white wavy markings on wings. Eggs are laid singly on leaves. Egg period 3 to 4 days. Larval duration will be completed within a period of 2 weeks. Pupation occurs among the leaf webbings and pupal duration lasts for 10 days. Several larvae can be seen inside each leaf web.

Cucumbers, muskmelons, watermelons, squashes, gourds, and pumpkins are commonly grown cucurbits in most parts of the world. Cucurbitaceous crops are the most important summer vegetables grown all over the world and it includes 118 Genera and 825 Species. Production and productivity of cucurbits are affected by the incidence of pests and diseases. Important pests attacking cucurbits are fruit fly, snake gourd semilooper, red pumkin beetles, hadda beetles/ epilachna beetles, pumkin caterpillar, vine borer/ clear winged moths, gall fly, serpentine leaf miner, shield bug, snake gourd bug and plume moth.
The melon fruit fly is distributed widely in temperate, tropical and subtropical regions of the world. It has been reported to damage 81 host plants and is a major pest of cucurbitaceous crops. The extent of loss varies between 30 –100 per cent depending on the cucurbit species and season. The adult is reddish brown with lemon yellow curved vertical markings on the thorax and fuscous shading on the outer margin of wings.
Biology: Eggs are white and cylindrical and laid singly or in clusters in cavities made with the ovipositor of the females on the fruits. The holes are sealed with gummy secretions, which solidify to form a shiny brown resinous material. Egg period is 1 to 2 days. The emerging maggots are apodous, dirty white, thicker at one end and tapering at the other and are seen within the fruits. Larval period is 7 days. The mature larvae emerge from the fruits and pupate in soil for a period of one week. Life cycle is completed in 15 to 20 days.

Cow pea is an important vegetable crop in Kerala. The crop was originated in Africa and is the major source of plant proteins in the diet of rural populations in Africa. The grains contain 25% protein and several vitamins and minerals. Insect pests pose the greatest threat to cowpea production. The crop is severely attacked at every stage of its growth by insects. High pest densities occur at many locations with complete loss of grain yield if no control measures are taken.
Moth has greyish brown forewings with large transverse hyaline band and a few smaller spots. Hind wings are hyaline with brown outer margin. The female moth lays eggs on or near flower buds. Caterpillars are brownish green with black warts. The young caterpillars bore into the buds, flowers or pods. Pods with bore holes may be seen. The infested flowers and pods may be webbed together. Flower drop may occur in severe cases. Caterpillars may bore into the stem also (Mahalakshmi et al., 2016).
Host range
Soybean, blackgram, greengram, Groundnut, common bean, lima bean, chickpea, mung bean, broad bean, pigeon pea.

Crucifers, also known as Brassicas or cole crops, are a group of vegetables belonging to the family Brassicaceae (formerly Cruciferae). Common cruciferous vegetables include cabbage, cauliflower, broccoli, kale, mustard, turnip, and radish. These crops are widely grown around the world due to their nutritional value, economic importance, and adaptability to diverse climates.
However, crucifers are highly susceptible to a wide range of insect pests that can cause significant yield losses if not managed properly. These pests attack various parts of the plant, including leaves, stems, roots, and heads, leading to both quantitative and qualitative damage.
Distribution
Crocidolomia binotalis is widely distributed in India and is particularly prevalent in regions where cruciferous vegetables are cultivated. The pest is active from the third week of November to the fourth week of December, with varying population densities depending on environmental conditions (Pawar et al., 2010)

Okra, commonly known as lady’s finger is a significant vegetable crops primarily grown in tropics and subtropics. Production and productivity of okra is affected by the incidence of pests and diseases. More than 70 insect pests have been recorded in okra (Ewete, 1978).
Biology: Forewings of the moth white with a pea- green wedge-shaped band running medially from base to outer margin of each wing. Eggs are laid on tender parts of the plant, leaf axils, bracts, leaf veins, buds, flowers and fruits singly or in two’s or threes. Egg period - 3-4 days. Caterpillars are brownish with median longitudinal streak dorsally and pale yellow or green ventrally. Larval period is 2 weeks. Pupae are seen on the plant in dirty white boat shaped cocoons made of silk. Pupal period is one week. The life cycle is completed in about a month.
Nature of damage and Symptoms
The newly hatched larvae bore into the terminal shoots or fruits. The damaged shoots droop, wither and dry up. The infested fruits are deformed with holes on them plugged with excreta.

Distribution
Thrips tabaci is present in the central and southern regions of India, occupying about 36.4% of the land area under the current climate conditions. In central India, the states of Maharashtra, Karnataka, Madhya Pradesh, Gujarat, and parts of Rajasthan offer highly suitable habitats. In the northern region, areas such as Uttar Pradesh, Bihar, Jharkhand, Chhattisgarh, Delhi, Haryana, Punjab, Uttarakhand, & parts of the northeastern states are favorable. In southern India, Karnataka, Andhra Pradesh, Telangana, & Tamil Nadu are particularly suitable.
Biology Adults: Adult onion thrips overwinter in soil in onion, small grain, and hay fields. More mobile than immature stages, adults can fly and are attracted to white and yellow colours, often landing on clothing or skin. They are elongated, with body colour ranging from yellow to brown, and have pale, fringed wings. Adults measure 1.0–1.3 mm for females and 0.7 mm for males. Their lifespan is 16–42 days on garlic and 28–30 days on onion. Females lay eggs for up to three weeks after a one-week preoviposition period. In spring, adults emerge, colonize weed hosts, & can fly or be carried by wind to new plants.
Eggs
Females lay eggs individually in leaf tissue, with one end near the surface for immatures to emerge. The eggs are microscopic, kidney-shaped, and initially white or yellow, turning orange with reddish eye spots as they mature. On onion, eggs average 0.23 mm in length and 0.08 mm in width. The incubation period is 4–5 days on onion, with hatching occurring in 2–3 days in the lab, but 5–10 days in cooler field conditions.

Brinjal (Solanum melongena), also known as eggplant or aubergine, is a major solanaceous vegetable crop grown in India and other parts of the world. It is affected by several insect pests throughout its growth period, significantly reducing both yield and quality.
Introduction
One of the major constraints in production and productivity of brinjal is the incidence of pests and diseases. The fruit and shoot borer is one of the most damaging pests of brinjal. Pest infestation starts from transplanting seedling stage and continues up to harvesting of the crop. Pest is active throughout the cropping season with peak infestation from last week of August to December
Distribution
China, India, Bangladesh, Malaysia, Thailand, Burma, Sri Lanka, Laos, Congo and South Africa

Chilli (Capsicum annuum L.), a member of the Solanaceae, is a significant spice and vegetable crop in India, extensively cultivated over warm temperate, tropical, and subtropical regions. Approximately 51 insect pests and two mite species from 27 families and 9 orders were discovered to be infesting chillies (Reddy and Puttaswamy, 1983). The yield losses range from 50-90 per cent due to insect pests in chilli (Nelson and Natrajan, 1994)
Chilli thrips: Scirtothrips dorsalis, Thripidae, Thysanoptera Biology
The adults are slender, yellowish brown in colour, with long, narrow, and heavily fringed wings. The adult female inserts eggs into the tissues of the leaves and shoots. The nymphs are similar to adults, except they are smaller and wingless. With the exception of the rainy season, the pest is present all year round. The life cycle is completed over a period of two weeks. Over the course of a year, multiple generations overlap.
Recently introduced invasive thrips in chilly is Thrips parvispinus Karny. It causes premature flower drop, shedding of fruits and malformations resulting in severe yield loss. The average fecundity is about 56 eggs with a mean developmental period of 18.8 days (Muray et al., 2009).

Fruit borer: Helicoverpa armigera, Noctuidae, Lepidoptera
Biology: The adult females are light pale brownish-yellow stout moths while the male moths are pale greenish with V-shaped speck. The eggs are creamy white and sculptured type. They are laid singly. The larva shows colour variation from greenish to brown. It has dark brown grey lines on the body with lateral white lines and also has a dark band. Pupation occurs in soil, leaf, fruit, and crop debris.
Nature of damage and symptoms
• The young larvae feed on tender foliage
• Mature larvae bore into fruit through circular holes
• Larva thrust only a part of its body into fruit and eat the inner content

Introduction
Potato is a tuber crop belonging to the Family solanaceae. In India large number of insects are attacking potato both in field as well as storage thereby causing both direct and indirect losses to farmers.
Distribution
Potatoes are cultivated in almost all states of India, but the majority of the crop is grown in the Indo-Gangetic plains of North India. The major potato growing states are Uttar Pradesh, West Bengal, Punjab, Bihar, and Karnataka (Rana and Anwer, 2018).
Distribution
Most potato cultivating fields of North India
Host plants: Carrot, broccoli, cabbage, mustard, radish, cucumber, pepper, wheat, rice (Holman, 2009).

Black pepper, popularly known as the “King of spices”, is the most popular spice crop in the world, and the crop originated in the Western Ghats and Malabar coast of India. India was the centre of pepper trade from time immemorial. Recently, black pepper production in the country remained stagnant at around 90000 tonnes in the last few years. Hence, India is losing its status as a leading producer and exporter of black pepper. Kerala accounts for 75 per cent of total black pepper production in the country. It has been noted that the increase in area under cultivation is not in tune with the increase in production. Low productivity is mainly due to the incidence of pests and diseases.
Black pepper is infested by 56 species of insects in India; among these, pollu beetle, (Lanka ramakrishnae Prathapan & Viraktamath), top shoot borer (Cydia hemidoxa Meyr), scale insects (Lepidosaphes piperis and Aspidiotus destructor) are considered as major pests (Devasahayam, 2000).
It is one of the most destructive pests of black pepper and the infestation is severe in plains of Malabar region (20-30 %) and low in hilly tracts (Wayanad and Idukki) and South Kerala.
Distribution: Tamil Nadu, Karnataka and Kerala.
Host plants: Castor, turmeric, guava, mulberry, etc.

Cardamom, often called the “Queen of Spices”, is one of the most valuable and aromatic spices in the world. It belongs to the family Zingiberaceae and the genus Elettaria. Cardamom, which is indigenous to South India's Western Ghats, is grown for its tiny, fragrant, green pods that contain tiny seeds that are used in cosmetics, medicine, and cooking. Kerala, Karnataka, and Tamil Nadu are the main cardamom-growing states in India, which ranks among the world's top producers. The crop grows best in shady regions with evenly distributed rainfall, rich forest soils, and humid, tropical weather.
Introduction
Shoot and capsule borer is a significant pest that infests a wide variety of wild and cultivated plants. Since larvae directly affect the reproductive plant parts, this pest becomes more significant in tropical and subtropical nations. Yellow peach moth, cardamom, or castor shoot and capsule borer are other names for the shoot and capsule borer (Thyagaraj, 2003).
Distribution: Tamil Nadu, Karnataka and Kerala. A serious pest of the nursery in cardamom
Host plants: Castor, turmeric, guava, mulberry, etc.

Cinnamon, commonly known is a highly valued spice crop obtained from the inner bark of the tree Cinnamomum verum, belonging to the family Lauraceae. It is native to Sri Lanka and Southern India, and widely cultivated in Kerala Tamil Nadu, and Karnataka. The spice is used globally in culinary, medicinal and aromatic industries. It thrives well in tropical humid climates with well-drained loamy soils and is generally propagated by seeds or cuttings. Though relatively hardy, cinnamon can be affected by a few key pests, especially when grown in monoculture or under poor management.
Biology: There are two adult forms clytia and dissimilis. The clytia are blackish brown wings with a series of arrowhead shaped white spots on the outer margin, and the dissimilis are black winged with elongated white spots and a series of marginal arrowhead shaped spots. Adults lay spherical, waxy looking, and orange-yellow colored egg singly on the upper and lower surfaces of young leaves. The emerging larva resembles bird droppings (defensive mimicry) at the initial stages. The fully-grown larvae are pale yellow with dark stripes on the sides. There are five larval instars, and the larval period lasts for 11–17 days. The elongated, brownish-black pupa is joined to the host plant’s stem by silky supports at the rear end. The pupal stage is 11–13 days long. The life cycle is completed in 24-36 days (Singh et al., 1978).

Distribution: India, China, Bangladesh, Sri Lanka, Taiwan, Thailand
Host: Turmeric, ginger, cardamom, castor, guava, mango, cocoa etc.
Biology: Adult moths are yellowish medium-sized moths. Wings are pale yellowish with black spots on the wings. The female moth lays round yellowish eggs singly or in groups on the tender part of the plant. Larva is long, pale greenish with a pinkish tint dorsally, head and pro-thoracic shield brown in color, body covered with minute hair. Pupation takes place in a loose silken cocoon inside the larval tunnel (Chong et al., 1991)
Nature of damage and symptoms
• Yellowing and drying of leaves of infested pseudostems
• Presence of a bore-hole on the pseudostem through which frass is extruded
• Withering and yellowing finally cause death of the central shoot
(dead heart) (Devasahayam et al., 2010 and Chong et al., 1991)
Management
• Pruning and destruction of infected plant parts
• Application of neem oil 1%

In India, jasmine is one of the oldest traditional flowers cultivated. It is infested by a number of insect and mite pests. The lepidopteran pests like bud borer, gallery worm, and leaf web-worm, are of significant importance. Sucking pests like eriophyid mite, lace wing bug, and thrips also causes minor damage to the crop. Dipteran pest like blossom midge also infests the crop. This chapter outlines brief descriptions of insect pests along with their hosts, life cycle, nature of damage, seasonal incidence and overall integrated pest management.
1. Bud worm: Hendecasis duplifascialis, Pyralidae, Lepidoptera
Budworm is an important pest of jasmine. Its damage ranged from 30 to 70 per cent, which had a significant impact on flower quality. The quality of opened flowers was affected by the silken tunnels and excretions created by budworm larvae inside the bud clusters (Gunasekaran, 1989 and Kamala and Kennedy, 2016).
The adult moths are small, pale white with wavy markings on wings and black patches on the wing margin. The moths lay eggs singly and glued on the unopened or immature buds, calyx and sometimes on the bud stalk, where the larvae eventually hatch and begin feeding.

Rose, one of the most popular cut flowers of the world known as the queen of flowers owing to its fragrance and aesthetic value. The rose has represented love, affection, innocence, and other noble traits for thousands of years. Roses are used for bouquets, flower arrangements, garlands, and worship. Apart from these, it is widely used in ayurvedic preparations, perfumes, soaps, cosmetics and also as flavouring agents in soft drinks and beverages.
1. Rose thrips: Rhipiphorothrips cruentatus, Thripidae, Thysanoptera
Adults are blackish brown in colour while nymphs are reddish in colour. Both adult and nymph lacerates leaves from the under surface and also on flower and flower buds.
Host range: Rose, grapes, Lagestoemia indica, Punica granatum.
Biology: Adult thrips lay about 45-55 eggs. Eggs are inserted into the plant tissues. The nymphal period is about 2-3 weeks, and the adult period is about five days.

Anthurium is a famous decorative cut flower and pot plant in the Araceae (order Spathiflorae) genus because of its extended vase life and beautiful, long-lasting inflorescences. After orchids, anthurium sales are the second-highest in the world.
Anthurium Thrips : Chaetanaphothrips orchidii Moulton
Order : Thysanoptera
Family : Thripidae
Biology: Chaetanaphothrips orchidii are small, pale yellow thrips commonly found in unopened anthurium buds and flowers. These pests are also commonly found in orchids. Females reproduce parthogenetically and on an average 80-100 eggs are laid by each female (Hata and Hara, 1998). Eggs are inserted inside the flower bud or leaf sheath. Nymphs are light yellowish in colour and are similar to the adults. Pupation takes place beneath the host plant in the soil or growing media. The entire life cycle lasts approximately 28 to 32 days but can extend up to 3 months, depending on the temperature. Increased temperatures and humidity, along with new growth of host plants, seem to promote thrips feeding and reproduction, resulting in heavier infestations and greater damage during the summer months

Orchids belong to the family Orchidaceae, one of the largest and most diverse families of flowering plants, with over 28,000 species and more than 100,000 hybrids worldwide. They are renowned for their exotic beauty, structural diversity, and ornamental value. Orchids, a diverse and prized group of ornamental plants, are sensitive to a variety of insect and mite pests. These pests affect orchids grown both under protected conditions (greenhouses, polyhouses) and in open environments.
Distribution: Kerala, Tamil Nadu, Karnataka, West Bengal, Assam, Sikkim, Maharashtra
Biology: Eggs are laid under the female’s shell and may remain after death. Crawlers are mobile and settle to feed, secreting protective coverings. Females remain stationary; males, where present, are short-lived and winged. Multiple overlapping generations per year occur under warm, humid conditions. Crawlers are the most susceptible to control treatments
(Smith et al., 2018).

Gladiolus is a genus comprising around 260 species of perennial flowering plants in the iris family (Iridaceae). Commonly known as "sword lilies," the name derives from their long, blade-like leaves—gladius being the Latin word for "sword." These plants grow from underground storage organs called corms and feature tall, unbranched stems that bear vibrant, trumpet-shaped flowers arranged in a single-sided spike. Their striking appearance makes them highly valued both in gardens and as cut flowers.
Most gladiolus species are native to sub-Saharan Africa, particularly South Africa, though some are also found in Europe and Asia. The flowers come in a wide variety of colors, forms, and often display intricate patterns or ruffled edges. Each flower is bisexual and sits between green bracts, with its tepals forming a characteristic funnel-shaped corolla. Known for their bold inflorescences and extended vase life, gladiolus flowers are a popular choice in floral design and landscaping worldwide

Chrysanthemum (Chrysanthemum spp.), popularly known as “mum” or “guldaudi”, is a major commercial ornamental plant grown for cut flowers, loose flowers, and garden display. Belonging to the family Asteraceae, it is valued for its vibrant colors, diverse flower shapes, and long vase life. Chrysanthemum is attacked by several insect and mite pests, affecting both foliage and floral quality, leading to reduced aesthetic and commercial value.
Distribution: Widely distributed in India wherever chrysanthemum and sunflower are grown.
Biology: The eggs are laid singly on the upper surface of leaves, obliquely inserted into the tissue, with opercula appearing as whitish or brown dots. The incubation period for the eggs lasts about 5 to 7 weeks. Following hatching, the nymphs undergo hemimetabolous development (no true pupal stage). The nymphal period spans 2 to 3 weeks, during which the nymphs moult five times before reaching adulthood. Adult insects are approximately 4 mm long, with blackish bodies and lace-like transparent wings (Rizkawati et al., 2023).

Carnation (Dianthus caryophyllus L.) belonging to the family Caryophyllaceae is very remarkable as a cut flower crop all over the world. It is well known for its attractive flowers with fragrance and varying colors. In India, the area under its cultivation is 2190 ha with a production of 8940 tones. But its production is affected by a number of insect pests and spider mite.
It is the two spotted spider mite or red spider mite which is considered as one of the most serious pests of carnation. High temperatures and low relative humidity favor the multiplication of these mites. These mites are found to be persisting throughout the year under polyhouse conditions.
Host range
Citrus, grapes, apples, pears, tomatoes, okra, potatoes, eggplant, beans, cucumbers, peppers, roses, jasmine, tea, coffee etc.

Lily (Lilium spp.) is a high-value bulbous ornamental plant prized for its large, attractive, and often fragrant flowers. Belonging to the family Liliaceae, lilies are widely grown for cut flowers, landscape decoration, and potted plant markets. Despite its beauty, lily cultivation is often challenged by several insect and mite pests that reduce aesthetic quality, market value, and bulb health.
Biology: Eggs are laid in clusters or singly on the undersides of lily leaves. They are yellowish to pale white, round, and smooth. Incubation lasts for about 3–6 days, depending on temperature and humidity. There are typically 5–6 larval instars. Newly hatched larvae are small and yellowish, later turning bright orange-red with black bands and spots. Caterpillars feed gregariously at first, skeletonizing leaves, then disperse to bore into stems or bulbs in later stages. The larval stage lasts for about 12–20 days. Pupation occurs in the soil, where larvae form a silken cocoon often mixed with soil particles. The pupal stage lasts 7–14 days. Pupae are reddish-brown and cylindrical. The adult is a medium-sized, metallic blue-black moth with bright yellow bands on the forewings and orange markings on the thorax. Adults are nocturnal and feed on nectar. Adult lifespan is around 4–6 days, during which female mate and lay eggs (Sathe, 2015).

Turfgrass refers to a group of grass species cultivated to form a dense, uniform ground cover, primarily used in lawns, golf courses, sports fields, parks, and landscape areas. Turfgrass serves both aesthetic and functional purposes, including erosion control, temperature regulation, and dust suppression. Turfgrass ecosystems provide a favorable habitat for various insect pests and mites that affect roots, stems, and leaves, compromising turf quality, appearance, and playability.
Distribution: Andhra Pradesh, Tamil Nadu, Maharashtra, Gujarat, Punjab, Uttar Pradesh, Odisha, and other regions, Mythimna separata (oriental armyworm), is more prevalent in the northern and eastern states.
Biology: Female moths lay eggs in clusters, often between leaf sheaths and blades, especially on dry grass. The eggs are initially pale yellow and darken before hatching. The larvae, also known as armyworms, go through several instars, feeding on grass blades.

Location and Layout
• As the laboratory will be handling materials hazardous to human and animal health, it must be isolated as much as possible from livestock yards, poultry farms and quarantine stations and should not function in or near establishments handling healthy animals, food products for human or animal consumption or near vaccine production laboratories.
• Animal houses, garages, workshops, stores, water tanks, residential accommodation, administrative blocks have to be constructed as per the need of ideal microbiological laboratory. Expansion possibilities should
also be kept in mind. The building can be arranged in 5 main groups:
• Administrative / laboratory rooms
• Decontamination and disposal area
• Animal house facilities
• Workshops / stores/garages and
• Residential accommodation for the staff, hospital, schools, playgrounds, parks, shops etc. • The laboratory building may be rectangular in blocks, arranged in L, T or U shapes and may be of one or several stories. The temperature, wind velocity, rainfall,

On the face of an outbreak of a disease, collection of right clinical specimens in proper preservative, their transportation to the laboratory under ideal conditions and quick processing of specimens in the laboratory to identify the causative agent are crucial to successful control of the disease. This attains more importance when a highly contagious disease appears.

Collection of specimens from a virus-infected host is the first step for demonstration or isolation of a virus. Specimens for virus isolation should be collected at the time when the maximum amount of virus can be expected to be present. i.e. collect the specimens as early as possible after the onset of clinical symptoms. In many viral diseases maximum viral production occurs before the patient becomes ill so that it is ideal to collect the specimen at an even early time.
The nature of the specimen taken in any particular instance is determined by the patient’s clinical syndrome and previous history. They are best considered under three main headings.

1. Specimen for virus isolation/antigen detection/ nucleic acid detection
2. Specimen for serological investigation
3. Specimen for direct examination

The chief functions of Clinical Microbiological laboratory are to examine and culture specimens for microorganisms, to make accurate species identification of important isolates and to perform antibiotic susceptibility tests when indicated. These tasks will assist physicians in the diagnosis and treatment of infectious diseases.

Once the specimen is received in the laboratory, the specimens are examined visually depending on the physicians order and the nature of the specimen. Wet mounts and smears may be prepared and stained for microscopic examination. Observations may or may not be immediately reported to the physicians depending on the definitiveness of the results. Timely information may often be used to establish a presumptive diagnosis and institute a specific course of therapy. Specimens that require definitive identification of potentially pathogenic microbes are processed further. All agar plates are streaked for colony isolation; then plates and broths. All cultures are placed in an incubator with appropriate temperature and environmental conditions to maximize the growth and replication of microbes. Often a presumptive microbial identificationcan be made. A final report should be delayed while subcultures and additional test procedures are performed to identify the organisms definitively.

Preliminary Identification of Unknown Bacteria
A number of techniques may be used in the direct microscopic examination of clinical specimens to demonstrate the presence of microbes The examination of wet mounts of unstained materials by phase contrast or
dark field microscopy is useful for demonstrating motility, spirochaetes and endospores. Giemsa, Wright or acridine orange stains may be helpful in observing bacterial forms that stain poorly or that have little contrast from
back ground material.

Special Staining
A. Spore stain
Spores may be spherical or oval and may be central subterminal or terminal in position within the cell. Shape, size and position of the spore in the cell are useful for the identification of sporulated organisms. Spores are difficult to stain and when once stained, they are difficult to decolourize. With Gram’s stain, mature spores remain unstained bodies. Modified Ziehl-Neelsen’s method of staining will be used for the staining of spores.
Composition
a. Concentrated carbol fuchsin
b. Sulphuric acid 0.5 % solution.
c. Loeffler’s alkaline methylene blue
Materials
Old cultures of B. subtilis and Cl. Chauvoei and staining solutions.
Procedure
1. Make smears and fix over flame.
2. Stain with carbol fuchsin and steam for 3 to 4 minutes.
3. Wash with tap water.
4. Decolourize with 0.5% sulphuric acid.
5. Wash with water.
6. Counter stain with Loeffler’s methylene blue for 2 minutes.

Antimicrobial Susceptibility Test
The routine use of the modified Kirby-Bauer method is recommended.
Material
• Sterile cotton swabs
• Soyabean – Casein digest broth or Tryptone soya broth
• Non selective agar such as nutrient agar.
• 0.5 Mc Farland turbidity standard:
Solution A
Bacl2, 2H2O. 1.75G
Make upto 100 ml with distilled water.
Solution B
H2SO4 1.0 ml
Make upto 100 ml with distilled water.
Stock Solution
0.5 ml of solution A + 99.5 ml of solution B. Dispense in 5 ml amounts.
Seal and store in dark.
• Glass vials or tubes 10 ml capacity
• Muller – Hinton agar
• Antimicrobial discs
• Sterile forceps
• Ruler or calipers

Viruses can be cultivated only in living cells. Laboratory animals, embryonated avian eggs and cell cultures are three systems available for this purpose. These three systems are used for the basic procedures of isolation and identification of viruses, maintenance of stock cultures and production of vaccines.
Cultivation of Viruses in Laboratory Animals
Animal inoculation helps to confirm the infectious nature of the disease. Laboratory animals used for many kinds of virological works such as clinical studies, pathogenicity of viruses, epizootiology, potency tests on vaccines and production of diagnostic and prophylactic antisera.

An ideal experimental animal is one that is
1. Uniformly and fully susceptible to the virus under study
2. Free from intercurrent infection and adventitious lesions and
3. Inexpensive purchase and maintain
If possible it is better to use germ free, specific pathogen free or colostrums deprived animals for experimental purpose. The same species of animals or atleast closely related species of the natural host is selected for inoculation, wherever possible. For example in case of swine fever, which shows a high degree of host specificity, the same species should be used for inoculation. in others such as in case of rabies with low host specificity, the causative virus will grow in a wide range of animals.

Molecular biology involves the study of DNA, RNA and proteins in cells. It emphasizes the biomolecules that carry out biological processes. One should understand the biomolecules participating in the process to understand the cellular function. In diagnostic microbiology, biomolecules are the analytes for testing and characterization. The analytes for molecular testing are the genomes (DNA), transcriptomes (RNA) and proteomes (Proteins) of the microorganism. Traditional methods of microorganism characterization depend on the culturing of the organisms, which is time-consuming and sometimes challenging to perform. Molecular techniques involve the characterization of DNA, RNA and protein to identify microorganisms. Nucleic acid sequence details are used to classify bacterial species, and mass spectrometry is used to identify microorganisms based on peptide profiles.

DNA (Deoxy ribonucleic acid)
DNA contains the genetic information required for the development and functioning of the living organism. It is the central informational molecule comprising a sequence of A, T, G and C nucelotides linked together by a phosphodiester bond. DNA is double helical in structure, composed of two complementary strands of nucleotides held together by hydrogen bonds between nitrogenous bases.

Precipitation and agglutination reactions are similar except in that for precipitation reaction antigen must be in soluble form. The precipitation reaction is highly specific and very sensitive for the detection of antigen being capable of demonstrating antigens in dilutions of 1:100,000 to 1: 1,000,000 or more. It is important to bear in mind that antigens used in precipitation reactions are of molecular size not cellular. A soluble antigen reacted with its homologous antibody at the optimum proportions produce an insoluble precipitate. The essential ingredients for precipitation test are soluble antigen (precipitinogen), antibody (precipitin) and an electrolyte (normal saline).
a) Precipitation ring test
The antigen is carefully layered over the antiserum, without mixing so that an interface is formed. Diffusion of each reagent will then occur into the other. If the system is homologous, precipitation will occur at the point in the tube, where the proper ratio of antigen to antibody is reached. Eg: Ascoli’s test for anthrax diagnosis.
In advanced decomposition of animals when cultures would be almost impossible, the precipitation method of Ascoli is used for diagnosis of Anthrax. This method is a mixing of immunized rabbit serum with an extract of organs under examination for the production of a precipitate.
Procedure
1. Grind up organs or blood of suspected animals.
2. Suspend in saline.
3. Boil for five minutes
4. Filter through filter paper and allow to cool
5. Carefully stratify 0.5 ml of this extract on 0.5 ml immune serum in small test tubes.

Among various immunodiagnostic tools enzyme immunoassay becomes increasingly popular for the detection of humoral immune response (seroconversion) or microbial antigens for the purpose of diagnosis. Enzyme Linked Immunosorbent Assay (ELISA) is widely used in studies of infectious diseases.Despite several merits of ELISA, it has some limitations such as requirement of ELISA reader to obtain accurate measurements. To overcome this, the dot immunobinding assay was developed .In Dot Immunoassay nitrocellulose membrane is used as an adsorbent matrix instead of plastic surface.The antigen is dotted onto the nitrocellulose membrane and the membrane then incubated with test antibody, and secondly with enzyme conjugated antibody directed against the first antibody.In case of positve
reaction, (specific antigen –antibody reaction) color dot is developed against the white background at the site of antigen deposition.In case of negative reaction, no color development takes place at the site of antigen deposition .
Principle
• In direct ELISA antibody is directly attached to solid phase and targeted by added antigen (detecting antigen).
• These added antigens are targeted by antispecies antibodies linked to enzyme termed as conjugates.
• The bound enzyme is developed by the addition of substrate /chromogen, then stopped and finally read using a spectrophotometer

Principle
The principle of the test is that antibodies possess two antigen-binding sites that will attach specifically with the antigenic determinants on the surfaces of the bacteria, cells or particles. Under suitable conditions one antibody molecule will combine with the determinant group on the surfaces of two bacteria and in this way a lattice is formed. These lattices are generally visible with the naked eye as clumps and they sediment readily due to the large size of the clump.
Pre-requisites for Agglutination Reaction
a) The antigen (agglutinogen) should be in particulate form, which will remain in suspension for sufficiently long time.
b) The antibody (agglutinin) should be directed to the target surface epitope.
c) Antigen – antibody should be present in optimal proportions.
d) An electrolyte is necessary for the reaction to take place and usually the reaction is carried out in physiological saline solutions (0.85% NaCl). Because of the overall negative charge on red cells or bacteria at neutral pH it is difficult for them to come close enough, so that for molecule such as IgG antibodies are able to form a bridge between epitope of two different cells or bacteria. The presence of salt tends to neutralize the charge effects to allow agglutination.
Plate Agglutination Test
This is employed to screen sera within a short period for preliminary testing.
Materials
• Brucella abortus coloured antigen
• suspected serum
• glass plate.