eChapters

Principle

The residue from the cell walls determination is extracted with boiling 8 percent sulphuric acid for one hour to hydrolyse the hemicellulose without dissolving cellulose, lignin or acid insoluble ash.

Equipment
    1.    Refluxing apparatus.
    2.     Filtering apparatus: same as described for cell wall determination
    3.    Filtering crucibles: same as described for cell wall determination.

Reagents

    1.    Sulphuric acid (8%) : Standardize Reagent grade sulphuric acid to specific gravity of 1.052 at 20ºC.
    2.    Bromocresol green indicator solution: Same as used for lignin determination.

(Method of Saryre, 1963 and IAEA, 1969)
Introduction

This technique helps in detection of very small amount of stable elements in fairly large quantitites of extraneous matter. Trace elements like zinc, copper, molybdenum and arsenic which exist in minute amounts in certain organs or tissues. For example, to detect the amount of copper present in one gram of tissue. using 1 mg of thin copper foil as control. Normal copper contains 69 percent ⁶³Cu and 31 percent ⁶⁵Cu. The (n,r) reactions on these two stable isotope produce ⁶⁴Cu and ⁶⁶Cu respectively. The half life of ⁶⁴Cu is 12.8 h and that of ⁶⁶Cu is five minutes. Therefore the latter can be ignored, if more than that an hour elapses between the end of the irradiation and the measurement. The specific activity of the unknown separate is compared with that of the known standard separate, and the content of Cu in the original tissue sample is thereby calculated.

(Method of Visek et al., 1953; Lueker and Lofgreen, 1961)

By using isotope dilution technique, It is possible to separate faecal calcium into endogenous and exogenous fractions. The true digestibility can be measured independently by making correction for faecal endogenous loss. The isotope dilution method described by Visek et al., (1953) involves daily intravenous injection of 45Ca for 15 days, followed by a collection of the serum and the faeces for 15 days. However, these numerous veinpunctures result in considerable discomfort to the animal. Lueker and Lofgreen (1961) modified this method in sheep by giving a single subcutaneous injection of 45Ca, followed by a collection of blood and the faeces for 7 days only.

The endogenous faecal calcium and true digestibility of calcium is calculated by the formula of Lofgreen and Kleiber (1954) as given below:

Introduction

It is a well known fact that the soluble oxalates are broken down by rumen microbes from buffalo and cattle and one of the end product formed is bicarbonates.

The rumen microbes are obtained from a fistulated buffalo calf fed on wheat bhusa and sarson cake supplemented with 60 g potassium oxalate daily for about three months. Representative sample of rumen ingesta is drawn before feeding and mixed culture is prepared by the method as described Donefer et al., (1960). Mixed culture and basal nutrient mixture (Quicke et al., 1959) of pH 6.9 mixed in equal quantities. Fifty millilitres of this mixture are added into each incubation flask containing one gram of wheat straw, ground to 40 mesh as substrate. In the incubation flasks are added 35,100 and 550 mg oxalic acid in the form of potassium oxalate and 0.24 µCi sodium oxalate 14C. Blank (containing no substrate, no oxalic acid added) and two controls, one having substrate but no added oxalic acid and the other containing the above plus radioactivity, are also run alongwith. All the incubation flasks are taken in duplicate. The flasks are incubated at 39ºC for 24 h  in an incubator and at the end of this period microbial activity is stopped by the addition of few drops of saturated solution of mercuric chloride.

Principle

    Inorganic salts are relativity insoluble in aromatic solvents which has been detrimental to the estimation of 45Calcium by scintillation counting. This technique inolves the counting of 45Calcium as calcium 2-ethyl hexanoate in a solution of toluene phosphor; and the counting as calcium chloride in a tertiary mixture of absolute ethyl alcohol-hydrochloric acid-toluene-phosphor.

Reagents
    1.    ⁴⁵Calcium. A ⁴⁵Calcium solution supplied as calcium chloride in hydrochloric acid solution, is suitably diluted.
    2.    Calcium chloride, 0.045 N, is prepared by dissolving ACS grade calcium carbonate, dried to constant weight at 110ºC, in 5 N hydrochloric acid.
    3.    Calcium acetate, citrate, lactate, acid phosphate, chloride and hydroxide.
    4.    Ammonium oxalate solution, 4 percent
    5.    2-Ethyl-hexanoic acid.
    6.    Toluene-diphenyloxazole solution, 2g of diphenyloxazole are dissolved in 500 ml of toluene.

(Method of Olbrich et al., 1971)

Jones and Ekman (1960) showed that ¹⁴⁴Ce is absorbed to a very small extent in the gut of the ruminant and suggested its use as a marker in digestibility trials. Huston and Ellis (1968) studied the affinity of radioactive cerium for feedstuffs and digesta particles in vitro experiments and found that cerium is rapidly absorbed onto and remained tenaciously bound to  digesta particles. Ellis and Huston (1968) tested radioactive cerium as a particulate flow marker in sheep with favourable results.

Experimental Procedure
Three Holstein bulls weighing 315, 239 and 305 kg and one Holstein steer weighing 327 kg are used. Twice daily, at 9.00 am and 9.00 pm, they are fed 3.4 kg of a balanced ration, meeting NRC requirements for normal growth and maintenance.

Separation of Bacteria and Protozoa
(Method of Czerkawski, 1975)

Rumen contents are obtained just before and two hours after feeding from sheep that had been given an experimental ration. The samples of rumen contents are strained through gauze and fractioned by sedimentation and centrifugation to give preparations of mixed protozoa (PR), large bacteria (LB) and small bacteria (SB). Whole particulate matter is also isolated by centrifugation at 20000g. The detail steps are mentioned in the flow chart given below.

(Method of Schram and Aines, 1959)

All the legume seeds and their byproducts are very rich in urease activity which may be responsible for an appreciable loss of urea nitrogen on long duration storage. Therefore it is necessary to estimate urease activity in legume seeds and their byproducts.

Reagents

    1.    Dimethyleaminobenzaldehyde solution (DMAB). Dissolve 16 g DMAB in 1 litre 95% ethyl alcohol and add 100 ml concentrated hydrochloric acid (Stable one month).
    2.    Pyrophosphate buffer. Dissolve 233 g Na4P2O7. 10H2O in approximately 980 ml distilled water. Add 3 ml of conc. Hcl and then dropwise further Hcl until the pH of the buffer is 7.7 to 7.8. Dilute to one litre.
    3.    Buffered urea solution. Dissolve 0.4 g urea in one litre pyrophosphate buffer (Stable one week).

Answer the followings
    Q.1.    What is the different stage of pathogenesis in mastitis?
    Ans.    There are three stages in pathogenesis 
These are as follows:
Invasion: here pathogens move from the exterior of teat to milk inside the teat canal.
Infection: Here pathogens multiply rapidly and invade the mammary gland.
Inflammation: In this stage clinical mastitis occurs with varying degree of clinical abnormalities of milk and udder.
    Q.2.    Of these phases, which phase is important to reduce the incidence of mastitis in control program?
    Ans.    Invasive stage is very important. The incidence of mastitis can be reduced by taking preventive measures at invasive stage by good management and good hygiene practices.
    Q.3.    What are the common abnormalities encountered in udder during mastitis?

Q.1.    What are the major clinical findings in mastitis?
    Ans.    These are abnormalities of secretions, abnormalities of udder related to size, consistency, temperature and sometimes systemic reactions.

Q.2.    How clinical forms of mastitis could be classified based of severity and duration?
    Ans.    Based on severity it is classified in four forms: Per acute mastitis, acute mastitis, subacute mastitis and subclinical mastitis. However, based on duration it is of three forms: Short term clinical or subclinical mastitis (Coliform mastitis), recurrent mastitis (Staphylococcus aureus mastitis, Streptococcal dysgalactiae mastitis) and persistent clinical or subclinical mastitis (Streptococcus agalactiae mastitis).

Q.3.    What is sub clinical mastitis?
    Ans.    In sub clinical mastitis there is evidence of inflammation which is evidenced by only raised SCC in milk without any visible abnormalities of milk and udder.

    Q.1.    How many species of Mycoplasma are responsible for causing mastitis?
    Ans.    At least seven species like Mycoplasma bovis, Mycoplasma Canadensis, Mycoplasma bovigenitalium, Mycoplasma alkalensis, Mycoplasma capricolium, Mycoplasma californicum, Mycoplasma disper and Ureaplasma sp. are responsible for causing mastitis

    Q.2.    Which herds are more prone to such type of mastitis?
    Ans.    The herds where milking hygiene is very poor and when the cows are imported from other farms or from public sale yards.

    Q.3.    Which organs act as common habitat of the Mycoplasma organisms?

    Q.1.    What are the most common isolated environmental Streptococci from mastitis cases?
    Ans.    Streptococcus uberis and Streptococcus dysgalactiae.

    Q.2.    What are the common sources of environmental Streptococcus uberis in a herd?
    Ans.    The common sources are bedding materials, lips, nostrils and abdominal skin of cows.

    Q.3.    What are the common sources of environmental Streptococcus dysgalactiae?
    Ans.    The common sources are tonsils, mouth, vagina and mammary gland of cows.

    Q.4.    Why incidence of Streptococcus uberis is highest just prior to calving?
    Ans.    The important reasons are the excessive accumulation of milk due to lack of milking, lack of keratin plugs from the streak canal and immunosuppression at periparturient period.

    Q.5.    The cattle fly sometimes involves in persistent bacterial contamination on teat of a cow. Mention the name of the cattle fly.

Fill in the blanks
    Q.1.    ………… organisms are the common isolates from clinically affected quarters.
    Ans.    Coagulase-negative staphylococci
    Q.2.    ………….is the most common cause of mastitis in goats.
    Ans.    Staphylococcus aureus
    Q.3.    Streptococcal zooepidermicus causes …………. in does.
    Ans.    Chronic suppurative mastitis.
 
Write True/ False about the statements
    Q.1.    Pseudomonas organism causes an acute mastitis characterized by extensive necrosis and fatal septicemia.
    Ans.    True.
    Q.2.    In summer mastitis, non-lactating goats develop severe mastitis whereas lactating goats suffer from moderate degree.
    Ans.    False

    Q.1.    How mastitis occurs in dogs? 
    Ans.    Mastitis in dogs initiates with the infection in the milk ducts of the dogs, where in the dog’s breast is enlarged as well as milk duct is blocked. The skin around the nipple might get cracked which might give a way to bacteria. In neglected cases it leads to cause gangrenous mastitis which is a severe form of mastitis in dogs.

    Q.2.    What are common pathogens responsible for mastitis in bitch?
    Ans.    Staphylococcus spp., Streptococcus spp. and Escherichia coli.
 
    Q.3.    What are the characteristic features of milk cytology during mastitis in bitch?
    Ans.    Milk cytology will show the presence of erythrocytes, degenerated neutrophils and bacterial agents.

    Q.4.    What characteristic physical changes of milk are seen during mastitis in bitch?
    Ans.    The viscosity of milk is increased and the colour of milk will be yellow to brown depending on the amount of blood and purulent exudates present.

Materials Required

Ostwald viscometer, stop watch, sample, distilled water, thermometer.

Procedure

    a.    Take sample and distilled water. Record their temperatures (they should be same for correct estimation). If the temperatures are not same bring both to a specific temperature say 20ºC or the room temperature.
    b.    Clean and dry the Ostwald viscometer properly before use
    c.    Fill the ostwald viscometer with distilled water and record the time taken (T₁) for the a known volume of water to flow through the orifice.
    d.    Empty the viscometer and dry it
    e.    Fill the ostwald viscometer with sample juice or liquid and record the time taken (T₂) for the a known volume of sample to flow through the orifice of the same viscometer.
    f.    Calculate the relative viscosity using the following formula

The students in batches will visit the laboratory of Plant Pathology for their acquaintance with different appliances, tools, glassware‘s and other miscellaneous items, which they will use for various exercises and experiments to be conducted. A list of some of them is given below:

Procedure

1.Peeled Potato slices are cooked in 500 ml of water by the time till they are easily penetrated by a glass rod.

2.Filtered with the help of muslin cloth.

3.Agar-agar is melted in another 500 ml of water.

4.Potato extract is added to the melted agar.

5.Dextrose is added in this mixture and stir well with the help of glass rod.

6.Make the volume to 1000 ml, by adding required water.

7.Medium is sterilized in an autoclave at 15 lbs pressure for 20 minutes at temperature of 121.6^oC.

Collection and dry preservation of disease samples

Materials : Iron knife, hand lens, vasculum, pruning scissors, paper envelops and plant press with blotters and news papers.

Method

•Collect infected plant parts and place them in paper envelops (use separate envelops for different kinds of diseased specimensxsxs).

•Place envelops in vasculum and bring to the lab.

•Arrange leaves and herbaceous stems inside the folder newspaper sheet.

•Place the newspaper between the blotters in plant press and tighten the bolts.

•After proper drying remove dried specimen from the press and keep them in separate folders.

•Label each folder giving the following informations.

a.Name of the organism

b.Habitat

c.Locality

d.Date of collection

e.Name of collector

f.Collection number

g.Other notes

Biological control of disease employs natural enemies of pests or pathogens to eradicate or control their population. Interactions that lead to biocontrol can include antibiosis, competition, induction of host resistance, and predation. Some of the microbial taxa that have been successfully commercialized and are currently marketed include bacteria belonging to the genera Agrobacterium, Bacillus, Pseudomonas and Streptomyces and fungi belonging to the genera Ampelomyces, Candida, Coniothyrium and Trichoderma.

•Dual Culture Technique: For in vitro screening, performance of antagonist isolates in dual culture (Morton and Stroube, 1955) with test pathogen can be tested.

•Pour 15-20 ml of sterilized melted PDA in 9 cm Petriplates and allowed to solidify.

•Place the bits (5 mm) of the test pathogen and antagonist on the PDA plates opposite to each other from 1.0 cm from the periphery of plates (if both are fast growing) or place it 2-3 cm apart (if both are slow growing).

•The Petriplates are incubated (at 28 ± 2^oC) for desired duration.

•Periodical observation on the growth of the antagonists and the ability of the antagonist to colonise the pathogen are recorded.

Q₁₀value: The number of times a reaction rate change with a 10^oC change in temperature is called Q ₁₀value. If reaction rate doubles with 10^oC change in temperature, Q ₁₀= 2. It is also called temperature quotient.

Q-enzyme: It is a branching enzyme responsible for formation of 1-6 bonds in amylopectin.

QHV process: It stands for Quality Has Value Process.

QO₂ : It is a unit for measurement of respiration. Or It is the oxygen uptake in microlitres per milligram on dry weight per hour.

Quailing: In this practice the oranges are kept in the baskets in which they are harvested to allow the fruit to loose little moisture. It is done for a couple of hours before transport to packhouse. This practice would allow the fruit to lose a little moisture which is called quailing. This operation is done for oranges to prevent Oleocellosis.

Qualitative measurement: It is a subjective / objective measurement of the quality of the product for presence of certain compound. It gives the idea of the presence or absence of the particular compound. It deals with the nature of the substance and not its amount or quantity.

Quality: Quality is a measure of the degree of excellence or degree of acceptability by the consumer. The quality may also be defined in term of end use and may vary depending upon consumer’s perception and need. Or quality is the totality of features and characteristics of a product that bear on its ability to satisfy stated or implied needs. It is a term defined by the consumer, buyer, grader, or any other client based on subjective and objective measurements of the food product.

Xanthan gum: It is a polysaccharide that is produced by fermentation of carbohydrates by a gramnegative bacterium (Xanthomonas campestris) and is a thickening and suspending agent used in pharmaceuticals and prepared foods. It is also called xanthan. Xanthan is a stabilizer produced by Xanthomonascompestris(Plate 257). Xanthan is not hydrolysed or degraded by humans.

Yabby: It is one of the burrowing Australian crayfishes (Cherax destructor) that are used for food.

Yakitori: A bite-size marinated pieces of beef, seafood, or chicken on skewers is called yakitori.

Yam: It is edible starchy tuberous root of plants (genus Dioscorea of the family Dioscoreaceae) used as a staple food in tropical areas.

Yautia: It is one of the aroid plants (Xanthosoma sagittifolium) chiefly of tropical America with starchy edible shaggy brown tubers that are cooked and eaten like yams or potatoes.

Yearling mutton: Meat obtained from 12-20 month aged sheep. It is also called Earling mutton.

Yeast: Unicellular plant body, very minute in size containing cell wall, cytoplasm with one or more vacuoles and a single nucleus, cause alcoholic fermentation (Plate 258). Yeast usually has little or no mycelium, and reproduces by budding. It results in production of yellowish surface froth or sediment that occurs in saccharine liquids (fruit juices) in which it promotes alcoholic fermentation, consists largely of cells of a fungus (as the saccharomyces, Saccharomyces cerevisiae), and is used in the making of alcoholic liquors and as a leaven in baking.

Any farmer/Entrepreneur who wishes to avail bank loan for starting or expanding his dairy enterprise should apply for loan to the nearest branch of a commercial bank, regional rural bank or co-operative bank in their area in the prescribed application form which is available in the branches of financing banks.

Preparing Projects for Bank Appraisal

    1.    The entrepreneur can prepare the project or Scheme report after consulting local technical persons of State Animal Husbandry Department, DRDA, Dairy Co-operative Society / Union / Federation / commercial dairy farmers.
 
    2.    The project report should have mention about practical training and experience in dairy farming. So in order to meet this requirement the inexperienced entrepreneurs can visit progressive dairy farms and government / military / agricultural university dairy farms in the vicinity to have first hand information about the enterprise. Nowadays several online portals (www.aaqaa.com.) also provide useful guidance and information on setting up dairy based enterprise.

1. Technical Feasibility of Dairy Unit

This includes following information such as
 
    1.    Availability of good quality animals in nearby dairy market,
 
    2.    Availability of good grazing grounds and lands,
 
    3.    Availability of green/dry fodder,
 
    4.    Availability of concentrate feed, medicines etc.
 
    5.    Further availability of veterinary aid, dairy breeding (AI) centre, milk collection and marketing centre are also important.

    6.    Nearness of the selected area to veterinary/breeding/milk collection centre and the financing bank’s branch.

    7.    Capability of the owner and employees such as technical knowledge and skill.

The Feasibility study of dairy project cover capital investment on land, building, infrastructure, working capital, manufacturing and marketing cost, and financial projections to work out its viability. Once viability is established, a detailed project report should include the following ten steps for project planning and implementation:

First Step: This step involves appointing a consultant to plan the project in relation to the design the dairy plant, proposed market and select an architect and a structural consultant. Prepare tender documents for civil works and plant equipment. Study the demand-supply of target products through a market survey to formulate the suitable product mix in terms of volume and range. e.g. study of various dairy products e.g. Khoa, ghee, Ice cream. The capacity of the plant and the provision of various facilities would be determined by the products demand.

• Introduction
• The Concept of a Farm Management System

Appendix A: Diagrammatic Presentation of Processes Involved in Obtaining an Indian Patent

For the diagnosis of clinical reproductive problems it is essential to collect blood, penile swab, Prepucial swab, prepucial washing, semen, smegma etc from breeding bulls. These samples are dispatched to the nearest laboratory for clinical diagnosis. 

5.1 Collection of Blood Serum

The bull is properly restrained and jugular vein area is cleaned and sterilized with 70% alcohol. An sterile hypodermic needle is used to collect 5-10 ml blood aseptically in a sterile test tube. This test tube is then kept in a slanting position. Serum is separated after the formation of blood clot and dispatched to the laboratory.

5.2 Collection of Seminal Plasma

Semen sample is collected as per the standard procedure. Carbolic acid or 1 % sodium acid is added to preserve seminal plasma at the rate of 2 drops in 1 ml ejaculate in the centrifuge tube. It is then centrifuged at 3000 rpm for 20 minutes. The supernatant is collected and stored in deep freeze till further analysis.

Bioinformatics is a multidisciplinary approach that combines computational and biological expertise to analyze biological data to advance research and development and it drew public attention with the announcement of the draft sequence of human genome by the then US President, Bill Clinton along with Craig Venter and Francis Collins in 2000. Bioinformatics and life sciences research are gaining attention in the government, industries and academic sectors. India was the first country in the world to establish a biotechnology information system (BTIS) network in 1987 through the Dept. of Biotechnology and created an infrastructure that enables it to harness biotechnology through the application of bioinformatics. Through this branch of science, biological data are acquired, stored and then analyzed. Bioinformatics can be used to store huge bioscience data for their future uses. By exploring computational tools, databases and new strategies for pests and diseases, resistant genes can be identified and plants with desired traits can be engineered to enhance agricultural productivity. Bioinformatics can be utilized in two ways for agriculture, viz., (i) at molecular level for application in breeding strategies and (ii) agri-informatics for land use planning, farmers window, early warning system, impact assessment and environmental applications.

The focus of world’s trade and economic activities is increasingly being shifted from resource-based economics to knowledge-based economics. This paradigm shift has been triggered due to the implementation of Convention on Biodiversity (CBD) and other related international treaties and policy frameworks. CBD for the first time states that bio-resources are the sovereign property of the countries in which they are found. It further stipulates that access to bio-resources by outside parties can be made only after prior approved consent of the concerned stakeholders and also with mutually agreed terms and conditions for equitable sharing of benefits accrued from the commercialization of biodiversity. To realize such proclaimed rights given in CBD and to fulfill obligations of various other relevant treaties, particularly in the context WTO regulation, it is imperative that the bio-resources-rich nations like India have comprehensive knowledgebase of the biodiversity and associated information systems in accessible form.

The advances in science and technology are deeply intertwined. Researchers can use the semiconductor manufacturing techniques that underlie miniaturization to build radios and exceptionally small mechanical structures that sense fields and forces in the physical world. These inexpensive, low-power communication devices can be deployed throughout a physical space, providing dense sensing close to physical phenomena, processing and communicating this information, and coordinating actions with other nodes. These multifunction devices are battery operated and are connected wirelessly with sensing, processing and communicating capabilities and are known as the wireless sensors. If we use these sensors with biological issues or Biotechnology, then we can find the new concepts biosensors. If we really implement these concepts in real life we can get rid of many diseases like heart attack, blood sugar level, gene reaction and other changes that happens in our body much before its shows to outside.

1.     Length and Size
2.     Volume
3.     Weight

Object: To demonstrate the phenomenon of osmosis using potato osmoscope. 

Material required: A potato tuber, sugar solution, beaker, water, petridish, knife, measuring cylinder, and stand.

Theory: Osmosis is a phenomenon in which water molecules diffuse through the semi permeable membrane from region of its higher concentration to the region of lower concentration.

Object: To demonstrate the phenomenon of endosmosis and exosmosis by using grapes and raisins.

Material required: Grapes, raisins, beaker or petridish, water and sugar solution.

Object: To demonstrate the process of diffusion.

Material required: Beakers, copper sulphate, distilled water and sugar solution.

Object: Estimation of water potential of plant tissue by Chardakov’s method.

Material required: Test tubes, sucrose or polyethylene glycol (PEG) 6000, methylene blue and dropper.

Principle: Concentration of the solution in which a plant tissue is immersed changes. If the solution is hypertonic (with lesser yw) the tissue loses water and the solution becomes dilute. If the solution is hypotonic (with higher yW) the tissue absorbs water and the Solution becomes concentrated. If the solution is isotonic (with yw equal to the tissue sap) the tissue neither loses nor absorbs water and the solution concentration does not change. Thus the yw of this solution gives the yw of the tissue.

Objective: Estimation of photosynthetic pigment from plant leaves.

Material required: Plant leaves, test tube, test tube stand, spectrophotometer, balance, centrifuge machine, mortar and pestle, acetone and distilled water.

Theory: Photosynthetic pigments, namely, the chlorophylls are present in green leaves and shoots of plants. There are at least five types of chlorophylls in plants. Chl’a’ and ‘b’ occur in higher plants, ferns and mosses. Chl ‘c’,‘d’ and ‘e’ are found only in algae and in certain bacteria. Chlorophylls are extracted from plant leaves. The absorption peaks of chl ‘a’ at 663 nm and ‘b’ at 645 nm are characteristic measures from which amount of chlorophylls can be calculated.

Object: To demonstrate chlorophyll fluorescence.

Material required: Spectrophotometer, fresh green spinach, mortar, pestle, acid washed sand, acetone, funnel, filter paper and test tubes.

Principle: Chlorophyll molecule absorbs light in the blue (450nm) and red (660nm) region and get into excited state and emits an electron. The electron drops back to the ground state within 10-9 seconds and some energy (which is not used in biochemical reaction) are given out as the observed fluorescent far red light.

Procedure and observation: Take some soft green spinach leaves. Grind these with a pestle -mortar in acetone adding a pinch of acid washed sand. Make a fine paste. Filter to get a clear solution in a test tube. Take the test tube in bright sun light.

Object: Measurement of leaf area by graph paper method.

Material required: leaves, graph paper and pencil or pen.

Object: Estimation of stomatal index and stomatal frequency with replica method.

Material required: Leaf sample, blue/red correction fluid, transparent finger nail polish, brush glass slide and microscope.

Principle: The number of stomata per unit area of the leaf is called as Stomatal Frequency. The stomatal index is defined as the percentage number of Stomata as compared to all the epidermal cells in a unit area of leaf i.e.

Object: Demonstration of leaf anatomy or structure of C₃ and C₄ Plants (demonstration by already prepared slides).

Material required: Already prepared slides of corn and wheat leaves and microscope.

Object: To demonstrate that water is lost from a living plant during transpiration. 

Material Required: A potted healthy plant, a bell jar, a rubber sheet or oil cloth, a glass sheet and grease.

Theory: Transpiration: Loss of water in the form of water vapour from the internal tissues of living plants through aerial parts of the plant is known as transpiration.

Object: To measure rate of transpiration of potted plant by gravimetric (Lysimeter) method. 

Material required: Potted plants, Polythene sheet, Platform and electric balance etc.

Method: Select same size and same type well watered potted intact plants. Seal the pots and the root system with a sheet of polythene so that the water loss to the atmosphere occurs only from the shoot system not from the soil (To check evaporation losses). Immediately record the weight of the potted plant and keep them in different environment such as (1) warm environment (2) cool environment (3) Calm environment.

Object: To measure the rate of transpiration by using Ganong’s potometer

Material required: Ganong’s potometer, beaker, wax or vaseline, stop watch and twig or a branch of a plant.

Theory: The amount of water absorbed is almost equal to the amount of water transpired. The difference between the two amounts is negligible. 

Method: Potometer consists of wide mouthed tube fitted with a hard cork in which a twig or branch of a plant is inserted. Other end of tube is dipped in beaker. One half of the tube is graduated. The apparatus is filled with water using the reservoir. An air bubble is inserted in the tube as shown in figure.

Object: To find out the relation between transpiration and absorption. 

Material Required: Wide mouthed glass bottle, with a narrow graduated side tube, split rubber cork, water and a plant with intact root system.

Theory: The loss in weight of plant indicates the amount of water transpired. And fall in water level of the side tube indicates the amount of water absorbed. This shows that amount of water transpired is normally equal to the amount of water absorbed by the plant roots.

Object: To demonstrate unequal transpiration by the two leaf surfaces from four leaf method. 

Material required: four leaves of china rose or any dorsiventral leaves, two stands, thread and grease or vaseline. 

Theory: Transpiration is a process in which loss of water occurs in the form of water vapour from the aerial parts of the living plant. 

Method: Take four leaves of equal size from a plant (e.g. China rose). Now apply or smear the vaseline on the four leaves as follows:

Object: To demonstrate that light and carbon dioxide are essential for photosynthesis (Moll’s half leaf experiment). 

Material Required: A potted plant, a wide mouthed bottle, caustic potash, splits of rubber Cork and vaseline etc. 

Object: To demonstrate that O₂ is evolved during photosynthesis. 
 

Material required: Hydrilla plant, beaker, test tube, funnel, pond water and pyrogallol. 
 

Theory: Oxygen is a byproduct of photosynthesis, which is evolved in presence of light and chlorophyll during photosynthesis.

Object: To demonstrate anaerobic respiration. 

Material required: Soaked pea or gram seeds, mercury, test tube stand, forceps and KOH pellets. 

Theory: Generally, plants at certain stage have the capacity to respire in absence of oxygen i.e. anaerobic respiration. In this process, oxidation of carbohydrate takes places, ethyl alcohol and CO2 being end products and only a little amount of energy is liberated-