eChapters

Materials Required 

Fruits/vegetables, hand refractometers, fruit pressure tester, weighing balance, scale, chemicals and glassware required for analysis.

Procedure

    1.    Go to the orchard or vegetable field and collect samples of fruits and vegetables
    2.    Apply the specific maturity indices to the samples

Materials Required

Fruits/vegetables, thermometer, refractometers, NaOH, Phenolphthalein, glassware, weighing balance, plastic crates for storage of commodity, water cooling facility.

Procedure

    1.    Harvest the fruit / vegetable
    2.    Count No. of unmarketable fruits in every 100 fruits harvested
    3.    Record the internal fruit temperature in about 5-10 randomly selected fruits with the help of a thermometer
    4.    Pre-cool the commodity in cold water (<5ºC), air or refrigerator till the temperature reaches near its optimum storage temperature
    5.    Record the internal fruit temperature
    6.    Record the time required to pre-cool the commodity
    7.    Keep the product for storage at ambient conditions alongwith control
    8.    Record the difference in the shelf life and quality during storage

Materials Required

Fruits/ vegetables, washing tanks/ containers, size grader, fruit waxes, packaging material, neem based fungicides etc. 

Procedure

    1.    Take available fruits or vegetables 
    2.    Wash them in plain water or chemical solution as told by the teacher
    3.    Shade dry the fruits / vegetables
    4.    Prepare suitable wax solution
    5.    Apply wax on to the fruits as told
    6.    Record the fruit weight before packing
    7.    Pack the fruits properly in the trays / boxes depending upon the type and purpose
    8.    Do curing if required 
    9.    Do pre-packaging, if desired
    10.    Label the package for desired information
    11.    Keep the fruits for storage
    12.    Record the observations pertaining to weight loss and rotting periodically depending upon the commodity and its shelf life.

Materials Required

Fruits/ vegetables, weighing balance, plastic crates, CFB boxes or other packaging material fruit pressure tester, hand refractometer, markers etc.

Procedure

    1.    Harvest the fruits
    2.    Subject them to various pre-treatments (as discussed in previous exercise).
    3.    Record their initial parameters i.e. weight, firmness, surface colour, TSS, rotting etc. 
    4.    Place properly packed commodity in storage alongwith a check under ambient conditions
    5.    Record the observations regarding changes in quality during storage
 

Materials Required

Fruit, pulper, fruit mill, hydraulic press, screw type juice extractor, stainless steel pans, chemical preservatives, glass bottles, closures, crown corks, crown corking machine, plastic barrels.

Procedure

    1.    Select and weigh sound fruits/vegetables for extraction of juice/pulp.    
    2.    Wash and peel the fruits as required.
    3.    Extract the juice by hot or cold process.
    4.    Pasteurize the juice/pulp to inactivate the enzymes and partially kill microorganisms.
    5.    Preserve the juice/pulp by heat processing after hot filling and hermetic sealing or by addition of permissible quantities of suitable preservatives.
    6.    Label the bottles properly and keep them for storage

Materials Required

Fruit juice/ pulp, cane sugar, potable water, essence, citric acid, colours, preservatives, weighing balance, bottling and sealing facilities, hand refractometer, homogenizer etc.  

Procedure

    1.    Extract the juice as discussed in previous exercise.
    2.    Filter the fresh or preserved juice through muslin cloth.
    3.    Calculate the quantities of juice, sugar, water, acid (if required), preservatives, colour, essence etc. required for preparation of a known quantity of                      squash as per FPO specifications.
    4.    Prepare the beverage as directed by instructor
    5.    Pack the beverage in bottles 
    6.    Seal the bottles and label them
    7.    Take out one of the bottles and measure its TSS and titratable acidity.

Materials Required

Fruits juice, enzymes (pectinase, cellulase, pectinase CCM etc.), filter press, buchner filter apparatus, water bath, filter paper pulp, buchhi type rotary vacuum evaporator, freeze concentrator, membrane concentrator (whichever available), vacuum grease.

Procedure

    1.    Extract the juice as discussed in earlier exercises. Take a known quantity of juice in a conical flask.
    2.    Add enzymes i.e. pectinase @ 0.2% to the juice and keep in a water bath at temperature of 40-50ºC for 2 hrs.
    3.    Shake the flasks occasionally for uniform mixing of enzyme all throughout the flask during the enzymatic treatment.
    4.    Leave the flasks undisturbed for 10-12 hrs for allowing the insoluble solids to settle at the bottom of the flask.
    5.    Siphon the clear juice from the top of the flask.
    6.    Pass this siphoned juice through filter press or through a 0.4-0.5 cm thick bed of filter paper pulp in a Buchner filter apparatus or filter press.
    7.    Record the volume of the clarified juice obtained. Calculate the wastage.
    8.    Concentrate the juice in rotary vacuum evaporator at 50+2ºC temperature and at 28-30” Hg vacuum to a TSS of about 50 to 70º Brix. Record the exact TSS.
    9.    Record the volume of condensate obtained in the round bottom flask of the buchhi apparatus.
    10.    Calculate the folds of concentration

Materials Required

Fruit pulp, cane sugar, potable water, essence, citric acid, colours, preservatives, weighing balance, glass jars (500 ml capacity), jar closures, hand refractometer, steel pan, ladle etc.

Procedure

    1.    Select sound and fresh fruits, free from any blemishes. Generally a mixture of ripe and unripe fruits should be taken as the ripe fruits impart good flavour and the unripe fruits are rich in pectin, required for a good set.

    2.    Wash the fruits thoroughly. Cut into 4-8 pieces depending upon the fruit and size
    3.    Boil the fruit pieces for about 20-25 min in boiling water and pass through pulper to get a good pulp.
    4.    Weigh known quantity of fresh or preserved pulp and add about equal quantity of sugar (preferably in 45:55 ratio)
    5.    Cook the mixture on gas flame by occasional stirring. 
    6.    When the mixture becomes thick, perform sheet test.
    7.    Citric acid is added generally towards the end of cooking. Addition of all the acid in the beginning may result in stickiness and poor set. 
    8.    Remove from the flame when the mixture gives a clear sheet
    9.    Flavours, colours etc may also be added towards the end of boiling. 
    10.    Fill the jam in glass jars (preferably pre-warmed in hot air).
    11.    Close the jars with lug caps and allow to cool overnight.
    12.    Label and store in a cool place.

Materials Required

Fruit extract, cane sugar, potable water, essence, citric acid, colours, preservatives, weighing balance, glass jars (500 ml capacity), jar closures, hand refractometer, steel pan, ladle etc.

Procedure for Jelly

    1.    Select sound and fresh fruits, free from any blemishes. Generally a mixture of ripe and unripe fruits shouldbe taken as the ripe fruits impart good flavour and the unripe fruits are rich in pectin, required for a good set.
    2.    Wash the fruits thoroughly. Cut into 4-8 pieces depending upon the fruit and size
    3.    Boil the fruit pieces in water (about ½ to an equal weight of fruits for fruits like apple and 2-3 times the weight of sliced fruits in case of citrus fruits) for about 30 min. with occasional stirring. Strain the boiled mixture through coarse muslin cloth to get a clear pectin extract
    4.    A second extract may also be taken by boiling the same mass again and straining.
    5.    Add sugar (usually 3/4th to one part of the extract or in 45:55 ratio) to the pectin extract and boil in a stainless steel pan. Add required quantities of acid towards the end of boiling.
    6.    Remove the scum rising on the top of boiling mass occasionally.
    7.    To avoid charring, occasional stirring is also done.
    8.    To avoid destruction of pectin the cooking should be completed fast
    9.    Determine the end point of jelly as detailed ahead.
    10.    Fill in hot dry glass jars and cool rapidly
    11.    Pour a thin layer of hot molten paraffin wax seal to avoid moulding.   
    12.    Label and store in a cool place.

Materials Required

Lime fruits, green chillies, ginger salt (common), stainless steel pans, kinives, weighing balance, spices etc. 

Procedure

    1.    Select sound and mature fruits of lime, mature but green chillies having good size and ginger rhizomes.
    2.    Wash the lime fruits, ginger and chillies properly in running cold water.
    3.    Cut the lime fruits into pieces / halves. 
    4.    Remove the stalks of the chillies and cut into longitudinal slices. Peel ginger and cut into long thin slices.
    5.    Put the lime pieces, chilli and ginger slices in a wide mouthed pre-sterilized glass jar 
    6.    Add common salt @ 250g / Kg of material.
    7.    Squeeze some of the juice over the slices
    8.    Stir the mass well mixing thoroughly
    9.    In case the juice does not cover the material completely, add pre-boiled and cooled 5% solution of citric acid.
    10.    Keep the jar in Sun for about a week with occasional shaking of the material.
    11.    When the lime pieces start turning light brown, the pickle is ready for consumption.

Materials Required

Fruits or fruit pulp, steel pans, weighing balance, refractometer, sugar, salt, spices, acetic acid, citric acid etc. 

Procedure

    1.    Extract fruit pulp as told in earlier exercises. 
    2.    Take a known quantity of freshly extracted or preserved pulp.
    3.    Weigh sugar (about half the weigh of pulp).
    4.    Heat pulp and sugar over a gas flame.
    5.    Add spices, chillies, salt, citric acid (if required) etc. and cook
    6.    Record the TSS to judge the end point. It should be more than 50o Brix.
    7.    Add measured quantities of acetic acid and sodium benzoate at the end.
    8.    Cool and pack in pre-washed and sterilized glass jars
    9.    Label and store.

Materials Required

Tomato fruits, pulper, steel pans, weighing balance, refractometer, sugar, salt, spices, acetic acid, citric acid, muslin cloth etc.

Procedure
I. Puree

    1.    Extract fruit pulp by Hot process as told by the teacher.
    2.    Record the TSS of the pulp.
    3.    Concentrate the pulp by heating to 12º Brix (heavy puree)
    4.    Hot fill the boiling puree into pre-sterilized glass bottles (200 ml cap.)
    5.    Seal the bottles by crown corking
    6.    Heat process in boiling water in a big aluminium pan for 15-20 min. 
    7.    Cool the bottles to room temperature as quickly as possible.
    8.    Label and store at a dry place.

Materials Required

Fruits or vegetables, stainless steel knife, steel pans, Can Body Reformer, Flanger, Double seamer, weighing balance, Refractometer / Salometer, sugar or salt etc. 

Procedure

    1.    Wash the selected fruit or vegetable (as the case may be) 
    2.    Cut into pieces or slices (the size and shape may depend on the fruit/ vegetable but it should not be too large or too small)
    3.    Blanch the material in boiling water for 1-2 min for effective inactivation of enzymes. 
    4.    Prepare the sugar syrup of strength 40o Brix or brine of 1-3 % strength.
    5.    Prepare the can body using a can body reformer and flange the ends in a flanger.
    6.    Seal the base plate using first and second roller operations of the double seamer. 
    7.    Transfer the prepared fruits / vegetables into the cans. 
    8.    Pour in hot syrup or brine into the cans.
    9.    Place the lid over the top of filled can. (follow the first roller operation of the double seamer, if the cans are OTS cans)
    10.    Exhaust the cans in boiling water for about 10 min. to exclude any dissolved air out of the tissues or syrup/brine.
    11.    When the temperature of the centre of can reaches 79-80 ºC for fruits or 82ºC for vegetables, remove the cans from the exhaust box and seal them properly (follow the second roller operation of the double seamer, if the cans are OTS can).
    12.    Process the sealed cans at 100ºC and 115-121ºC for fruits and vegetables respectively for 15-30 min in case of fruits and 25-45 min. in case of vegetables.
    13.    Cool the cans immediately in cold water and air dry them at room temperature.
    14.    Label and store in a cool dry place.

Materials Required

Canned product, weighing balance, can cutter, vacuum/pressure guage, seam checking guage, Refractometer / Salometer etc. 

Procedure

    1.    Take a canned product say canned peach.
    2.    Record the weight of the can (A)
    3.    Record the information marked on the label i.e net  weight, date of packing, etc
    4.    Observe the can externally for the concavity of the lid.
    5.    Record the vacuum of the can by using a vacuum guage
    6.    Cut the lid of the can with a can cutter
    7.    Record the head space
    8.    Put all the contents over a sieve and let the liquid drain
    9.    Record the weight of empty can (B) to calculate net contents
    10.    Record the drained weight of the pieces.
    11.    Record whether the liquid is syrup or brine
    12.    Record the TSS of the salometer reading of the liquid (as the case may be)

Materials Required

Dehydrator, balance, thermometer, sulphur dioxide, blanching pan, fruits / vegetables, polythene or aluminium bags.

Procedure

    1.    Select sound fruits or vegetables 
    2.    Record their initial weight and wash them properly in plain water to remove any dirt or extraneous matter. 
    3.    Peel with stainless steel knife (as the case may be) and cut into thin uniform pieces or slices of  0.2-0.5 mm thickness
    4.    Blanch the slices in boiling water for 1-2 min. to inactivate the enzymes followed by immediate cooling.
    5.    Treat the slices with sulphur (sulphuring or sulphiting) in order to prevent darkening of the product during drying. 
    6.    Dry the slices in a mechanical air dehydrator at about 55 + 5oC temperature.
    7.    Record the weight of the product after every 2 hr till it becomes (almost) constant i.e. when there no more (significant) loss of moisture from the product.
    8.    Pack the product in ploythene or aluminium laminated pouches and seal them air tight. 
    9.    If it is to be converted into powder, the dried product can be ground in a grinder before packing.
    10.    Label the packages and store them in a cool dry place.
    11.    Plot the drying rate curve of the product on a graph paper and analyse it.
    12.    Record the initial and final moisture content of the product (as discussed in Exercise No. 19). 

Materials Required

Apple (or any local fruit), fruit mill, hydraulic press, weighing balance, Refractometer/sugar pectinol, diammonium hydrogen phosphate, Saccharomyces cerevisiae culture, Potassium metabisulphite (KMS) etc.

Procedure

    1.    Take a known quantity of apple (or any other fruit) say 20 Kg
    2.    Extract the juice as discussed in earlier exercises.
    3.    Add about 200 ppm of kms.
    4.    Add sugar syrup to increase the TSS to about 20 – 24ºBrix.
    5.    To about 10 Kg of juice diammonium hydrogen phosphate (10 g) and pectinol (20-25 g are added).
    6.    Take about 500 ml of this mixture in a conical flask and add pure Saccharomyces cerevisiae culture @ 1 % and plug the neck with cotton. Keep at 22-24ºC to complete the fermentation within one day and yeast culture is ready.
    7.    Add this culture to about 10 Kg of must for further fermentation (i.e. culture is added @ 5% therefore 500 g is added to 10 Kg must).
    8.    Record the TSS of the must before fermentation.
    9.    Keep must for fermentation in big jars (narrow mouth) filled to about 75 % of their volume. Close the mouth of jars or bottles with cotton plug.
    10.    Allow fermentation to take place at 22-24ºC for about 10–15 days.
    11.    When no more bubbles are coming out of the jars, fermentation is complete. At this stage the TSS reaches about 7º Brix.
    12.    Siphon the wine. Add filter aids if wine is not clear.
    13.    Blend the wine with sweetening agents if required.
    14.    Fill in beer bottles by keeping a head space of about 2.5 cm followed by crown corking them.
    15.    Pasteurize the bottles by keeping at 62.5ºC for 20 min.

Materials Required

Cut flowers, 1000 ml conical flasks, scissor, tape, distilled water, chemicals, weighing balance, volumetric flasks, labels, thermometer (max. & min. type) and hygrometer. 

Procedure

    1.    Collect fresh spikes of flowers
    2.    Cut at the base keeping about 25-60 cm of the cut flowers stick (uniformly for each flower).
    3.    Prepare Pulsing/ Vase solutions.
    4.    Put the prepared flower spikes in labeled conical flasks containing different Pulsing/ Vase solutions (for a known period of time).
    5.    Record the weight of the spikes and other observations daily.
    6.    Record the temperature and relative humidity of the storage chamber daily.
    7.    Analyse the recorded data and draw conclusion about the best vase solutions for the selected cut flower.

Materials Required

Fruit, vegetable or product sample, hot air oven, weighing balance, crucibles, petri plates, lead pencil / marker etc.

Procedure

    1.    Take crucibles  or petri plates and record the tare weight of crucibles / petri plates (C) and mark them
    2.    Cut the sample into small and thin pieces (if solid)
    3.    Put the sample into the crucibles / petri plates in three replications
    4.    Record the weight of crucibles + sample (C + S)
    5.    Put in hot air oven for drying at 70 + 2ºC.
    6.    Record the loss in weight after every 1-2 hrs till it becomes constant
    7.    Calculate the moisture and total solids. 

Materials Required

Sample, hot air oven, weighing balance, crucibles, muffle furnace, lead pencil / marker etc.

Procedure

    1.    After following the procedure for the determination of moisture (in silica crucibles as given in previous chapter) the same sample can be used for the determination of ash. 
    2.    Heat the dried sample in the silica crucibles over a heater/hot plate/burner to burn the carbon.
    3.    When it becomes completely black and no more smoke comes out of the sample, shift them to muffle furnace.
    4.    After heating the samples in muffle furnace, all the marks of pen / pencil/ marker are lost, so one would not be able to identify the samples and error of weights may occur. Therefore, it is advised to place the samples in the furnace in one or two rows and draw the diagram on a piece of paper for identification purpose.

Materials Required

Fruit/ sample, refractometers, thermometers, standard solutions of known TSS, temperature correction Tables.

Procedure

    1.    Calibrate refractometers of all ranges to be used.
    2.    The TSS of the juices/pulps/fresh fruits can be measured with the one having lowest range i.e. 0-32º Brix while, for measuring the TSS of concentrates, syrups, squashes, jams, jellies etc. refractometers of the higher ranges are made use of.
    3.    Put a drop of material on to the glass surface of refrectrometer’s prism, cover gently with the lid and observe the TSS against light.
    4.    Measure the temperature of the liquid which is being evaluated and apply temperature corrections (Annexure III).
    5.    Record the corrected value as the TSS of the sample.

Materials Required

Sample, 0.1 N NaOH solution, 1% phenolphthalein indicator solution, beakers, conical flasks, pestle and mortar, funnels, volumetric flasks, pipettes, pH meter, shaker.

Reagents

    1.    0.1 N NaOH: Dissolve 4 g NaOH (AR) crystals in sufficient water to make up the final vol. 1 litre.

    2.    1 % Phenolphthalein: Dissolve 1 g in 100 ml water. Readymade indicator solutions are more commonly used.  

Procedure

Preparation of sample: For fresh fruits and vegetables, prepare a well blended, uniform pulp in a pestle and mortar and for processed products, take out a representative sample and blend it well so that it becomes uniformly soluble when added to water. Filter through a fine muslin cloth or coarse filter paper if felt necessary. The liquid samples can be taken directly for estimation.
    1.    Take out a known weight / vol. (say 20 g) of sample in a beaker (in 3 replicates).
    2.    Make it to a known vol. (say 100 ml) with added distilled water.
    3.    Take 10 ml of this aliquot in a conical flask.
    4.    Add 1-2 drops of phenolphthalein indicator into the aliquot.
    5.    Titrate against 0.1 N solution of NaOH till light pink coloured end point is obtained.
    6.    For coloured products, i.e. tomato products where judgment of light pink colour is difficult, titration may be done over a shaker, and digital pH meter may be used for continuously measuring pH of aliquot which would show a steady increase with more and more addition of 0.1 N NaOH. Titre value may be recorded when pH of 8.3 is obtained.
    7.    Record the titre value and calculate the acidity of the sample as per the given formula.

Materials Required

Sample, reagents, beakers, conical flasks, pestle and mortar, funnels, volumetric flasks, pipettes, hot plate, weighing balance, spectrophotometer/colorimeter.

Reagents

    1.    0.1 N NaOH: Dissolve 4 g NaOH (AR) crystals in sufficient water to make up the final volume 1 Litre.
    2.    1 % Phenolphthalein: Dissolve 1 g in 100 ml water. Readymade indicator solutions are more commonly used.
    3.    Alkaline copper Tartarate
        a.    Dissolve 12.5 g anhydrous sodium carbonate, 10 g sodium bicarbonate, 12.5 g potassium sodium tartarate and 100 g anhydrous sodium sulphate in about 400 ml of distilled water and make up the final volume to 500 ml.
        b.    Dissolve 15 g copper sulphate in distilled water. Add one drop of sulphuric acid and make up to 100 ml in distilled water.
    4.    Always prepare fresh Alkaline Copper Tartarate reagent by mixing 96 ml of sol. a. and 4 ml of sol. b. and before use.
    5.     Arsenomolybdate Reagent: Dissolve 5.0 g ammonium molybdate (AR) in 90 ml of distilled water. Add about 5.0 ml of concentrated sulphuric acid and mix well. Add 600 mg of disodium hydrogen arsenate dissolved in 50 ml water. Mix well and incubate the prepared reagent at 37ºC for 24-48 hr or 55ºC for 30  min. with constant stirring.

Materials Required

Sample, reagents, beakers, conical flasks, pestle and mortar, funnels, volumetric flasks, pipettes, weighing balance, spectrophotometer/colorimeter.

Reagents

    1.    2% Metaphosphoric Acid Solution: Weigh 20 g of HPO₃ and dilute to 1 litre in water or 4% oxalic acid.
    2.    Dye Solution: Dissolve 100 mg of 2, 6 Dichlorophenol-indophenol dye (Phenolindo-2,6- dichlorophenol sodium salt, AR, C₁₂H₆Cl₂NNaO₂ 2-3H₂O) and 84 mg of sodium bicarbonate (sodium hydrogen carbonate AR, NaHCO₃) in hot (85-95ºC) distilled water. Cool and make up the vol 100 ml. Filter and dilute 25 ml of the dye to 500 ml with distilled water. 
    3.    Standard Ascorbic Acid: Weigh 50 mg of ascorbic acid (AR) and make up to 50 ml with 2% metaphosphoric acid or 4% Oxalic acid solution (1000 ppm i.e. 1000 µg/ml). Dilute 4 ml of this solution to 100 ml with 2% HPO₃ or 4% oxalic acid. (1ml = 40 µg ascorbic acid).

Materials Required

Specific gravity bottles, weighing balance, sample, pipettes, thermometer, distilled water 

Procedure

    1.    Take the sample and distilled water record their temperatures (they should be same for correct estimation). If the temperatures are not same bring both to 20ºC or the room temperature.
    2.    Clean and dry the specific gravity bottles properly before use.
    3.    Record the tare weight of the specific gravity bottle (T) alongwith the stopper.
    4.    Carefully fill the specific gravity bottle with distilled water up to the brim and then place the stopper tightly so as to exclude extra water. 
    5.    Dry the exteriors of the bottle with a tissue paper
    6.    Record the weight of (bottle + water) say A
    7.    Similarly, fill the same specific gravity bottle with test sample and record the weight (bottle + sample) say B
    8.    Calculate the specific gravity of solution using the following formula

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

Materials Required

Spectrophotometer, ethyl alcohol, beakers, pestle and mortar etc. 

Procedure

    1.     Preparation of sample extracts
        a.    Liquids : Centrifuge the juice at 5000 rpm for 4-5 min., and collect the supernatant. For liquid samples prepare a 40 % solution in ethyl alcohol. To 30 ml of the sample add 45 ml of ethyl alcohol and mix thoroughly. Filter through Watman 
No. 1 paper.

        b.    Semi-solid samples: To 15 g of blended sample add 15 ml of distilled water and 45 ml of ethyl alcohol. Mix thoroughly and filter through Watman No. 1 paper.
        c.    Dried products: Extract known quantity of sample (4-6 g) with 100 ml of 60% ethyl alcohol for 12 hrs and filter through Watman No. 1 filter paper.
    2.     Measure the absorbance of the samples at 440 nm for non-enzymatic browning and 400 or 420 nm for measurement of colour.
    3.     Prepare 60% ethyl alcohol solution as blank. Add 30 ml of distilled water to 45 ml of absolute alcohol and use as blank.

Materials Required

Sample, Liebig condenser, digestion flasks (700-750 ml cap), conical flasks, filtering cloth (thick linen cloth for clear filtration) reagents, muffle furnace, crucibles.

Reagents

    1.    0.255 N Sulphuric acid solution : Dilute 2.5 g (1.36 ml) of concentrated H₂SO₄ to 200 ml. Ensure correct concentration by titration.
    2.    0.313 N Sodium hydroxide solution: 2.504 g NaOH / 200 ml water nearly free from sodium carbonate. Ensure correct concentration by titration.
    3.    Asbestos Chips: gooch grade, medium fibre, acid wash and ignited
    4.    10% Potassium sulphate (K₂SO₄) solution: Dissolve 10 g of Potassium Sulphate (AR) in water and make up to 100 ml.

Materials Required

Soxhlet apparatus, petroleum ether, water bath, sample, grinder, weighing balance, filter paper, threads.

Procedure

    1.    Take the given sample and weigh it (A)
    2.    Dry the samples in an oven till there is no more weight loss and weight it (B)
    3.    Grind the sample into powder.
    4.    Place a weighed quantity of sample in filter paper, wrap the paper and tie it tightly with a thread so that no powder comes out of the paper during extraction. 
    5.    Pour approximately 100 ml of anhydrous ether into the bottom flask. Turn the heaters on. Set the temperature at about 40ºC.  
    6.    The ether will evaporate and circulate through the sample and again will come back to the flask. 
    7.    Repeat the process for about 16 – 18 hrs for complete extraction. 
    8.    Remove the sample and evaporate most of the ether from the mixture of crude fat and ether obtained during extraction.
    9.    Evaporate excess of ether from the crude fat sample by evaporation on a water bath. Evaporate till it is free from the fumes of ether. Record the weight (C).

Materials Required

Sensory scoring card, products, pen/ pencil for coding, plain water

Procedure

    1.    Assign codes to the samples i.e. a, b, c, …… or 1, 2, 3, ….. etc.
    2.    Keep the coded samples randomly (eg. c, f, a, b, d, e) in a row for evaluation
    3.    Start the evaluation of samples for various sensory attributes one by one. 
    4.    Evaluator should rinse his/ her mouth after each sample so as to avoid overlapping of taste, flavour etc.
    5.    Assign marks from 1-9 based on pleasurable and un pleasurable experiences.
    6.    Compute the means for a particular attribute on the basis of the evaluation of different evaluators (replications).
    7.    Find out the best sample on the basis of highest average scores obtained.

Materials Required

Sensory scoring card, products, pen/pencil for coding, plain water.

Procedure

    1.    Take the given two samples
    2.    Make them three by duplicating/repeating either one of the two
    3.    Assign codes to the samples eg. AAB, ABA, ABB,  BAB etc.
    4.    Ask the evaluators to evaluate the samples for any differences between the 3 on the given Specimen Evaluation Card.
    5.    Find out how many of the evaluators have chosen the odd sample correctly (say it is 16 out of 20 evaluators).
    6.    Now find out the degree of difference in the samples indicated by the evaluators who correctly found the odd sample (Say it is much as per 3, moderate as per 8 and slight as per 5 out of the 16 correct evaluators.).
    7.    Find out how many of the correct evaluators preferred A sample and how many the B sample (say 11 preferred A sample and 5 B sample).
    8.    After going through Annexure II you may conclude that the two samples differed significantly from each other as 16 out of 20 evaluators found the odd sample correct. But, the preference was not significant as it should have been at least 13 out of 16 judges indicating same preference so as to make it significant. So, it may be concluded that the two samples differ significantly but both are acceptable.

Materials Required

Sensory scoring card, products, pen/ pencil for coding, plain water.

Procedure

    1.    Assign codes to the samples i.e. a, b, c, …… or 1, 2, 3, ….. etc.
    2.    Keep the coded samples randomly (eg. c, f, a, b, d, e) in a row for evaluation
    3.    Start the evaluation of samples one by one. 
    4.    Evaluator should rank the samples on the basis of intensity of a single sensory attribute according to his / her preference.
    5.    Compute the means on the basis of the evaluation of different evaluators (replications).
    6.    List the samples in increasing or decreasing order of preference.

Materials Required

Record book

Procedure

    1.    Follow the instructions given by the teacher and the factory representative.
    2.    Listen carefully to the explanations of the factory representative
    3.    Observe the functioning of various equipments
    4.    Ask queries wherever not understood properly.
    5.    Note down the observations in your record book  

Observations

    1.    Name and address of the factory or processing unit
    2.    Size of the Unit (Large, Small, Cottage, Home)
    3.    FPO Licence No.    
    4.    List of products manufactured    
    5.    Height of the processing plant    
    6.    Information regarding raw material store, finished product storage    
    7.    List of equipments installed in the factory
    8.    Water supply system     
    9.    Waste disposal system    
    10.    Hygiene in the surroundings    
    11.    Any other

Synonyms: Hydrophobia, Lyssa, Jalatanka

This is an acute highly fatal viral disease of all warm blooded animals including man characterized by signs of abnormal behaviour, nervous symptoms (increased excitability and irritability), impairment of consciousness ascending paralysis and death. The disease was variously known as lytta or lyssa coming from the belief that the disease was caused by a worm under the tongue (lytta) or hydrophobia, which describes the thirst and fear of water associated with the disease. The English name rabies derived from Latin word ‘Rabere’ which has been derived from Sanskrit word ‘Rabhas’ meaning raging, furious, savage, madness or to do violence whereas Greek term hydrophobia is now specifically used for rabies in man. The symptom hydrophobia (fear of water) is developed due to painful spasm of muscles of deglutition.

Distribution: The disease is known since ancient time. The disease has been recorded in various parts of the world in different species of animals. Australia, Britain, Scandinavian, New Zealand, Singapore, Hong Kong, Hawaii and some other Islands are free from rabies. This was possible due to strict quarantine measures and restriction on import of animal and animal products from disease prone areas. Lot of outbreaks are reported in India throughout the year causing 30,000 death annually and number of death toll may be higher as many cases go unnoticed. India accounts for the 80% casualties from rabies worldwide.

Avian influenza (AI) is a listed disease of the World Organization for Animal Health (OIE) that has become a disease of great concern for both animal and human health. Avian influenza (AI) or 'bird flu' is an infectious disease of birds ranging from a mild to a severe form of illness. It is caused by influenza A virus under the family Orthomyxoviridae. It is an enveloped virus with two types of surface projections haemagglutinin (H) and neuraminidase (N). The haemagglutinin is a rod shaped homotrimer which agglutinates RBCs and neuraminidase is a mushroom shaped homotetramer which releases the virus from cells because of its sialidase activity. There are 17 different H antigens (H1 to H17) and 10 neuraminidase antigens (N1 to N10). The virus also contains 8 single stranded negative sense RNA segments each of which encodes for a protein. Avian influenza virus is generally limited to poultry but an unprecedented epizootic of avian influenza A (H5N1) that is highly pathogenic has crossed the species barrier in Asia to cause many human fatalities and likely to possess an increasing pandemic threat. Zoonotic potential of AI virus was first recorded in 1997 with hospitalization of 18 people (six died) following the infection with HPAI (H5N1) in Hong Kong. This provided the evidence that HPAI strains of chicken can adapt, multiply and even cause death in affected individuals. So, it is the prime time to create awareness about the HPAI and its transmission to human beings so that necessary precautionary measures can be taken to prevent and control the disease efficiently and effectively.

Etiology: Influenza viruses are under the Orthomyxoviridae family and belong to either influenza A or influenza B or influenza C genus. The virus particles are enveloped with glycoprotein projections having haemagglutinin or neuraminidase activity and contain 8 negative sense single stranded RNA. Influenza A viruses are classified into subtypes based on these two surface antigens, haemagglutinin (H) and neuraminidase (N ). The haemagglutinin is a rod shaped trimer which agglutinates RBCs and neuraminidase is a mushroom shaped tetramer which releases the virus from cells because of its sialidase activity. Beneath the lipid envelope there is a membrane or matrix (M) protein which surrounds a helical complex containing the nucleoprotein (NP) and polymerases (PB1, PB2, P) protein. There are also two nonstructural protein (NS) present in the virus. There are 17 haemagglutinin antigen (H1 to H17) and 10 neuraminidase antigens (N1 to N10) present in influenza A viruses. Each RNA strand of AI virus code for at least one protein. The H glycoprotein of influenza virus has two important functions which are imperative for the infectivity of the virus (i) it is responsible for attachment of the virus to the host cell receptors of glycoprotein or gangliosides containing sialic acid. (ii) it possesses fusion activity between the host cell membrane and the virus membrane so that virus genetic material is introduced into the host cell. The glycoprotein is produced as a precursor, HAo which requires post translational cleavage to HA1 and HA2, by host proteases before it is able to induce membrane fusion. Then only the B virus particles becomes infectious. Type A viruses include the avian, swine, equine and human influenza A viruses. The large number of subtypes present in the type A influenza viruses are due to fact that viruses often undergo antigenic shift by the process of genetic reassortment.

Swine influenza (Swineflu) is a highly contagious viral disease of pigs. It is an upper respiratory tract disease of pigs that causes outbreaks in herds. Swineflu infects people every year and is found typically in people who have been in contact with pigs although there have been person to person transmission of the disease. Influenza A virus subtypes namely H1N1, H1N2 and H3N2 are circulating around the world. In US, H1N1 subtype was exclusively prevalent among swine population before 1998 however since late August 1998, H3N2 subtype has been isolated from pigs. Most of the H3N2 virus isolates are triple assortants containing genes from human (HA, NA and PB1), swine (NS, NP and M) and avian (PB2 and PA) lineages. Influenza viruses are generally species specific i.e. avian influenza virus will infect only birds, swine influenza virus pigs, human influenza virus humans and so on. Although interspecies transmission of influenza viruses are reported, the transmission of influenza virus from one species to another is not always possible. Again, influenza A virus isolated from pigs can acquire the pandemic potential but not isolated from other species. Further, recent outbreaks of swineflu affecting human beings in Mexico, USA, UK,Canada, Asia etc with a death toll of 49 persons resulted in high alert in all the countries and compulsory checking of all the persons coming from the affected countries to other counties. The causative agent of swineflu is H1N1. So, it is the prime time to create awareness about the disease among the public which in turn help in tackling the disease in an effective and efficient manner and to prevent its transmission to human beings from swine species.

Etiology: Swineflu is caused by a virus under the genus Influenza A virus and family Orthomyxoviridae. Influenza A viruses are spherical with a diameter of 80-120 nm and may be filamentous. There are two types of surface glycoproteins namely haemagglutinin (HA) and neuraminidase (NA) present on the virus particles. A total of 17HA and 10NA types have been reported in influenza A viruses. It contains 8 copies single stranded negative sense RNA molecules as genome which encode 12 proteins: 3 subunits of the viral RNA polymerase complex, PB1 (encoding 3 proteins PB1, PB1-F2 and N-40), PB2 and PA; a nucleoprotein (NP), two surface glycoproteins, HA and NA, a matrix protein (M1), an ion channel protein (M2), a nonstructural protein (NS1) and a nuclear export protein (NEP- also known as NS2). There are 170 combinations of two surface glycoproteins, 17 HA and 10 NA that determine the antigenic properties and subtype classification of virus. However, human disease historically has been caused by 3 subtypes of HA (H1, H2 and H3) and two types of N (N1 and N2). Further in 1997, the outbreak of avianflu/birdflu in Hong Kong affected 18 persons with a death of 6 persons. The avian influenza virus responsible for that outbreak was H5N1. From that time H5, H7 and H9 ( all from avian origin) have been implicated in causing disease in human beings.

Dengue is a mosquito-borne viral disease and considered to be a major health problem globally. It is also called break-bone fever, dandy fever, denguero bouquet fever, giraffe fever, Polka fever and 5 days or 7 days fever. There are two important manifestations, namely dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS) which are responsible for heavy morbidity and mortality every year causing serious concern to health. It is an important disease caused by an arbovirus worldwide especially in the tropical and subtropical regions. The severity of the disease can be judged from simple statistics that over 40 per cent (2.5 billion) of the world's population in 100 countries continue to live under threat of contracting dengue infection. Annually, dengue virus infections are responsible for up to 100 million cases of dengue fever and more than 500,000 cases of haemorrhagic fever. Besides, every year the disease forces nearly 500,000 people to hospitalization of which 90% are children and causing 24,000 deaths. Mostly these deaths are due to lack of early diagnosis of dengue virus infection. In DHF case fatality rate generally exceeds 20 per cent in a non-endemic population and it can be reduced to less than 1 per cent with the aid of modern supportive therapy based on early diagnosis of the type of viral infection.

Dengue fever (DF) is an acute febrile viral disease characterized by headaches, bone or joint and muscular pains, rash and leucopenia as symptoms. Dengue haemorrhagic fever (DHF) is characterized by four major clinical manifestations: high fever, haemorrhagic phenomena, 73 often with hepatomegaly and in severe cases, signs of circulatory failure. Such patients may develop hypovolaemic shock resulting from plasma leakage. This is called dengue shock syndrome (DSS) and can be fatal.

The year 2001 witnessed unprecedented global dengue epidemic activity in the American hemisphere, the Pacific Islands and continental Asia. During 2002, more than 30 Latin American countries reported over 1 million DF cases with large number of DHF cases. This has been followed by extensive epidemics of DHF in several parts of India during 2003.

Dengue fever (DF) is caused by ingestion of viraemic blood containing dengue virus by mosquitoes of Aedes sp. followed by passage to a second human host. An extrinsic incubation period is required before the virus appears in the saliva of the transmitting female Aedes mosquitoes. After the bite, there is an incubation period of 3 to 14 days (average 4 to 7 days) followed by the onset of fever and nonspecific signs and symptoms (frontal headache, retro-orbital pain, body aches, nausea and vomiting, joint pains, weakness and rash). The febrile phase is self limiting and may last for 2 to 10 days followed by remission of fever in most cases. Laboratory abnormalities include neutropenia followed by a lymphocytosis and thrombocytopenia.

Chikungunya has been emerged as an important public health problem in the Indian Ocean Islands and India recently. Epidemiological, geographical and demographic data suggested that 1.38 million people were affected by the disease in India in 2006. The actual incidence of the disease may have been higher but underreporting did not reveal the actual picture. A large proportion of those who became ill consulted private health care professional or sought alternative medical care such as Ayurveda or homeopathy not included in the data collected by Govt. officials.

During the epidemic, rural hospitals, private and public clinics flooded with victims of Chikungunya. The virus did not kill its victims but inflicted considerable pain and misery besides local, regional and national financial burden towards health care. Only National Institute of Virology (NIV) and National Institute of Communicable Disease (NICD) provided accurate diagnosis of the disease. The Chikungunya assumed epidemic proportions because viral agent mutated, the vectors discovered the new ways to spread and the host lacked immunity to fight the disease. Modern transport systems - cars, buses, trains and aircraft particularly the movement of humans played a key role in spreading the disease that helped to maintain man-mosquito man cycle.

Etiology: First isolated in Tanzania in 1953, Chikungunya is under genus Alphavirus and family Togaviridae. It contains a single stranded positive sense RNA with a length of 12 Kb coding 4 non-structural (nspl - nsp 4) and 3 structural proteins (E1-E3). Among the structural protein, El protein correlates with serological response in human hosts and also modulates penetration of the virus in the mosquito species. Genetic analyses and historical accounts suggest that the Chikungunya virus originated in tropical Africa and subsequently evolved into 3 distinct genotypes - the East African, the West African and the Asian genotypes. The Asian genotypes have a high degree of nucleic acid sequence homology among themselves but the African strains exhibit wider sequence diversity and have been shown to undergo genetic micro-evolutions even during the course of an epidemic. Prior to the 2006 epidemic in India, the 3 genotypes were restricted to the geographical areas only. Previous outbreaks in India (1963 and 1973) were caused by the Asian genotypes, but the 2005 epidemic in the Indian Ocean Islands and the 2006 epidemic in India have been attributed to the East African genotype which arrived in India 5 years ago.

This is an acute viral encephalitis having public health importance. It is under the genus Flavivirus and family Flaviviridae. The genus Flavivirus comprises of group of arbovirus though a few members of the genus have no known vector. The genus contains over 70 viruses of which 40 are mosquito borne, 16 are tick borne and 18 have no known vector. The type species of the genus is yellow fever virus. Although all flaviviruses are serologically relate, they can also be grouped serologically into distinct groups: dengue serological group, the JE serological group and a less serologically cohesive yellow fever virus group. The disease has been recognized in Japan since 1871 and was named Japanese B encephalitis to distinguish it from 'encephalitis A' (encephalitis lethargica, Von economo's disease) which was then prevalent. The virus was first isolated in Japan during an epidemic in 1935.

Etiology: JE virus is under the genus Flavivirus and family Flaviviridae. The genome is about 11 kb which encodes 3 structural proteins (C, capsid protein, PrM the membrane precursor protein that is cleaved proteolytically by a cellular protease to form the M protein in mature virions, and E, envelope protein) and 7 non-structural (NS) protein (NS1, NS2a, NS2b, NS3, NS4a, NS4b, NS5). The Eglycoprotein is the most immunologically important protein. The Japanese encephalitis serological group of flaviviruses comprises eight virus species and two subtype viruses with a geographic range encompassing all continents except Antarctica. The major virus species and their geographic range are as follows: the Japanese encephalitis virus ( JEV) in eastern, southern and southeastern Asia, Papua New Guinea and the Torres Strait of northern Australia; the West Nile virus (WNV) in Africa, southern and central Europe, India, the Middle East and North America and as Kunjin virus ( a subtype of WNV) in Australia and Papua New Guinea; the Murray Valley encephalitis virus (MVEV) in Australia, Papua New Guinea and the western Indonesian archipelago and the St. Louis encephalitis virus (SLEV) in North and South America. The other minor members of the group are Usutu (USUV), Koutango and Yaounde viruses in Africa; Cacipacore virus in South America and Alfuy, a subtype of MVEV, in Australia.

Hendra, a disease of horse has been recognized in Australia since 1994-95 with zoonotic importance. It is caused by a virus under the genus Megamyxovirus and subfamily Paramyxovirinae and family Paramyxoviridae. This virus is also endemic in certain species of bat. Another zoonotic bat virus called Nipah virus has been placed under the same genus and family. It is transmitted between the horses through contact. Infected horses can develop a severe and fatal respiratory disease characterized by dyspnoea, vascular endothelial damage, pulmonary oedema and neurological signs. Experimental infection of cats and guinea pigs showed fatal respiratory disease. The transmission of virus from horse to horse and horse to man is through close contact. One person died of the disease due to respiratory failure and another one due to delayed onset of encephalitis. A number of diagnostic methods are available to provide prompt, sensitive and accurate diagnosis of the disease.

History: In 1994-95, a new viral zoonotic disease was identified in Queensland, Australia resulting in death of 2 human and 15 horses in two separate incidents. Again, in 1999 another horse died at the same place. The term 'Hendra' has been derived from the name of a place, suburb of Brisbane where first outbreak took place. In September 1994, a total of 15 horses died due to hyperacute respiratory disease. The trainer of the horse also died. In 1995, a farmer died of meningoencephalitis after having contact with a Hendra positive horse. In the 1999, a mare died of Hendra virus infections.

Etiology: The Hendra virus is under the new genus Megamyxovirus and family Paramyxoviridae. The Hendra virus was isolated in Vero cells from lung specimens and produced CPE within 3 days. The virus can grow in MDCK, BHK, RK13, LLC-MK2 and MRC5 cells. It also grows in cells derived from birds, reptiles, amphibians, fish and in embryonated chicken eggs. The virus does not have haemagglutinin or neuraminidase activity. The virus is pleomorphic in shape, 38-600 nm in diameter and enveloped with two glycosylated transmembrane proteins of different lengths (15 and 18 nm). It differs from respirovirus and rubulavirus in not using sialic acid containing receptors. The virions are labile and infectivity is destroyed by heat, lipid solvents, non-ionic detergents, formaldehyde and oxidizing agents. Virions contain one molecule of negative sense ssRNA about 15 Kb long which codes 8 proteins. The Hendra virus is the largest of all known viruses under Paramyxoviridae. The genetic studies showed that Hendra virus is more similar to Morbillivirus or intermediate between Morbillivirus and Respirovirus. The amino acid homology between Hendravirus and Morbillivirus varies from 17-21% (P protein) to 40-42% (M protein) compared to 6-10% and 8% respectively for other members of Paramyxoviridae. A new Hendra like virus has been identified as a cause of respiratory and neurological disease in pigs and humans in Malaysia, called Nipah virus, after a village where the first human case occurred. (The amino acid homology of N protein between Hendra and Nipah is 92%, whereas in case of P&V it is 80-85%). Like, Hendra virus, the Nipah virus is thought to have emerged from a fruit bat (Pteropus hypomelanus).

Nipah virus infection is an emerging zoonotic disease first described in 1999 and the virus is under the genus Henipavirus and family Paramyxoviridae. The disease is similar to another emerging zoonotic disease caused by Hendra virus and can spread from its wildlife reservoir to pigs and humans where it causes an often fatal disease. It is a notifiable disease in most countries and to be handled under highest biological security level 4 (BSL-4). In Malayasia an outbreak of viral encephalitis occurred over a 35 week period from September 29, 1998 to 31^st May, 1999 among pig farmers creating considerable anxiety and fear. Of the 265 cases reported to the Ministry of Health in Malaysia, 105 were fatal and the case fatality rate was 39.6%. The outbreak also spread to neighbouring Singapore where 11 workers of an abattoir developed the disease with one fatality. 93% of the reported cases involved those directly in the pig farming industries namely pig farm workers, pig farm proprietors, housewives and family members assisting in the farms. The majority of the cases occurred in those aged 20 and above (92.4%) and actively engaged in pig industry followed by the age group 10-19 years (6.4%). About 82.6% of the cases are male.

Etiology: Henipavirus is a genus of the family Paramyxoviridae and order Mononegavirales containing two members, Hendravirus and Nipah virus. The henipaviruses are naturally harboured by Pteropid fruit bats (flying foxes) and are characterized by a large genome (18.2 Kb), wide host range and their recent emergence as zoonotic pathogens capable of causing illness and death in domestic animals and humans.

The viruses are pleomorphic ranging in size from 40-600 nm in diameter. They possess a lipid membrane overlying a shell of viral matrix protein. At the core is a single helical strand of genomic RNA tightly bound to N (nucleocapsid) protein and associated with the L (large) and P (phospoprotein) proteins which provides the RNA polymerase activity. F (fusion) protein trimers and G (attachment) protein tetramers are embedded within the lipid membrane. The genome of the Nipah virus consists of a non-segmented, single stranded negative sense RNA. In common with other members of the Paramyxovirinae subfamily, the number of nucleotides in the Henipavirus genome is a multiple of six, known as the 'rule of six'. The Nipah virus has been classified by the CDC as a Category C agent.

Hantaviruses have been responsible for a variety of human illnesses particularly haemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). There are currently 47 recognized sero/genotypes of hantaviruses with considerable geographic diversity. Each hantavirus type appears to be specific to a different rodent host. The different hantavirus types are associated with different types of disease both in terms of target organs and disease severity. HFRS is primarily a disease of the Old World while HPS is a disease of New World. Although the disease may have first been recognized as early as 1000 years ago in China, it affected >3000 United Nations Soldiers during Korean war (1950-1953). Now it is reported that hantavirus affect as many as 2 lakhs people each year in Asia and Europe. The agent of this virus remained unknown until 1978 when Lee and his associates isolated and named Hantaan virus, after a river in South Korea. Other viruses antigenically related to Hantaan virus were characterized and placed under the genus Hantavirus and family Bunyaviridae.

Over the past few decades, the sharp increase of incidences of hantavirus disease in humans throughout the world has been reported. There are 47 recognized hantaviruses with considerable geographic diversity. But the number of hantavirus virus types recognized continues to grow. Hantaviruses cause persistent infections in rodent reservoirs which directly impact transmission of these zoonotic agents between rodents and from rodents to humans. Environmental, 127 demographic and physiological factors likely interact to mediate responses to hantaviruses in natural rodent populations. Although not much work has been carried out, comparisons of human and rodent immune responses to hantaviruses may provide insights into why hantaviruses often are fatal for humans but cause only persistent infections in rodents.

Marburg virus (MV) disease was named after the West German town where the largest outbreaks of human infection was reported. It is also known as Vervet monkey or green monkey disease as monkey species introduced the disease in Europe. Marburg disease virus was reported in 1967 in Frankfurt-on-Main and Marburg, West Germany and Belgrade, Serbia in vaccine factory workers who came in contact with blood, organs or cell cultures of African green monkeys. Ebola and Marburg hemorrhagic fevers are severe zoonotic diseases seen not only in humans but also in non-human primates. Most species of Ebolavirus and the only known species of Marburg virus occur in Africa. Primates are infected sporadically from an unknown source probably bats. Humans seem to become infected directly from bats in caves, as well as when they, come in contact with tissues from infected apes and other species. Once the virus has entered the population, it can spread from person to person. Some epidemics affect hundreds of people. Although the mortality rate varies, 90% case fatalities have been reported in the most pathogenic viruses. No vaccine is available and the only treatment is supportive. Epizootics in gorillas and chimpanzees are equally serious, and appear to threaten the survival of these species in the wild. Other wild mammals including duikers also seem to be killed during outbreaks. One species of Ebolavirus, Reston Ebolavirus, occurs in the Philippines. Until 2008, this virus was known only as an infection of nonhuman primates. It does not appear to be a human pathogen; although some people in contact with this virus have seroconverted, they remained asymptomatic. In December 2008, Reston ebolavirus was discovered in pigs during an unusually severe outbreak of porcine reproductive and respiratory syndrome (PRRS) in the Philippines.

Etiology: Ebola and Marburg hemorrhagic fever are caused by members of the genera Ebolavirus and Marburgvirus, respectively. These viruses are the only members of the family Filoviridae. The genus Ebolavirus contains four recognized species: Zaire ebolavirus (formerly Zaire Ebola virus), Sudan ebolavirus (formerly Sudan Ebola virus), Ivory Coast ebolavirus (formerly Cote d'Ivoire Ebola virus) and Reston ebolavirus (formerly Reston Ebola virus). At least 13 strains of these viruses have been identified. A fifth species, tentatively named Bundibugyo ebolavirus, was isolated from a recent outbreak in Uganda. Marburg virus contains a single species, Lake Victoria Marburgvirus (formerly Marburg virus). Six strains of Lake Victoria Marburgvirus had been recognized as of 1990; at least nine genetically distinct strains were identified from a more recent outbreak in the Democratic Republic of the Congo. Marburg virus contains ssRNA as genome. The virion is highly pleomorphic and measures 80 nm in diameter and 700-1000 nm in length. Filoviruses can be destroyed by autoclaving. Treatment with sodium hypochlorite (1:100 dilution of household bleach) or standard hospital disinfectants can also be used. Ebolaviruses are reported to be susceptible to 2% sodium hypochlorite, 2% glutaraldehyde, 5% peracetic acid and 1% formalin. These viruses can also be inactivated by ultraviolet light, gamma irradiation, 0.3% betapropiolactone for 30 minutes at 37°C (98.6°F), or heating to 60°C (140°F) for 1 hour. Marburgvirus is susceptible to 1% sodium hypochlorite, 2% glutaraldehyde or formaldehyde, ultraviolet light and heat.

Crimean Congo hemorrhagic fever (CCHF) was first reported in the 12^th century as a hemorrhagic syndrome in present day Tajikistan. However, in the modern era, the first outbreak of the CCHF was reported in 1944-1945 in the Crimea region with more than 200 cases and the disease was called Crimean Hemorrhagic Fever. Ten years later in 1956, the virus was isolated from a febrile patient in Belgian Congo and this isolate was noted to have the same antigenic structure with the Crimean strains. For this reason, the virus was called Crimean Congo Hemorrhagic Fever. Nowadays, outbreaks of CCHF have been documented in Africa, the Middle East, Eastern Europe, and Western Asia including India. CCHF is a severe hemorrhagic fever with a case fatality rate of up to 50%.

Etiology: It is a tick-borne viral disease belonging to the family Bunyaviridae, genus Nairovirus. Like other Nairoviruses, CCHF virus is an enveloped single stranded negative-sense RNA virus and its tripartite genome consists of a small (S), a medium (M) and large (L) segment which encode for the nucleocapsid protein (NP), the envelope glycoproteins G1 and G2 and an RNA-dependent polymerase, respectively. The virus is transmitted to humans through the bite of infected ticks or by direct contact with viremic animals or humans. Infected humans can spread the disease via close contacts which may result in community outbreaks and nosocomial infections. It is an important human pathogen and could be used as a bioterrorism agent.

Pathogenesis: The pathogenesis of CCHF is not well understood. A common pathogenic feature of hemorrhagic fever viruses is their ability to disable the host immune response by attacking and manipulating the cells that initiate the antiviral response. This damage is characterized by marked replication of the virus together with dysregulation of the vascular system and lymphoid organs. Endothelial damage contributes to hemostatic failure by stimulating platelet aggregation and degranulation, with consequent activation of the intrinsic coagulation cascade. DIC is noted as an early and prominent feature of the disease process. In another study, the levels of interleukin IL-1, IL-6, and TNF-alpha were higher among patients who subsequently died as compared to survivors. The DIC score was higher among fatal cases, correlating positively with IL-6 and TNF alpha levels, and negatively with IL-10 levels. These findings demonstrate that proinflammatory cytokines play a major role in pathogenesis and mortality of patients with CCHF.

Synonyms: Splenic fever, wool sorter's disease, Tarka

It is an infectious disease of livestock and worldwide in distribution. This is a zoonotic disease. It is also known as splenic fever because of enlargement of the spleen (splenomegaly).

Etiology: It is caused by a rod shaped bacteria of 4-8 µm X 1-1.2 µm known as Bacillus anthracis. It is non-motile, capsulated, aerobic, spore bearing and Gm+ in nature. The organisms remain in chains. It was the first pathogenic bacterium to be observed under the microscope, the first communicable disease to be transmitted by inoculation of infected blood, the first bacillus to be isolated in pure culture and shown to possess spores and the first bacterium used for preparation of an attenuated vaccine.

The capsule is polypeptide in nature being composed of a polymer of d(-) glutamic acid. Capsules are not formed under ordinary conditions of culture but only if the media contain added bicarbonate or are incubated under 10-25% CO₂. Capsules are formed in the body of an infected animal.

Spores are formed in culture or in the soil, but never in the animal body during life. Spores are formed only in the presence of O₂. Spores are central, elliptical or oval in shape. The spores are very resistant to physical and chemical agents. They have been isolated from naturally infected soil for as long as 60 years. They resist dry heat at 140°C for 1-3 h and boiling for 10 min.

Anthrax bacilli is Gm + and non-acid fast. When blood films containing anthrax bacilli are stained with polychrome methylene blue for a few seconds and examined under the microscope, an amorphous purplish material (capsule) is noticed around the bacilli called M'Fadyean's reaction. On agar places, irregularly round colonies are formed 2-3 mm in diameter raised, dull opaque, grayish white with a frosted glass appearance. Under the low power microscope, the edge of the colony is composed of long, interlacing chain of bacilli, resembling locks of matted hair. This is called "Medusa head appearance." Virulent capsulated strains form rough colonies while avirulent or attenuated strains form smooth colonies. On gelatin stab culture, a characteristic 'inverted fir tree' appearance is seen with slow liquefaction commencing from the top.

Synonyms: Pearl's disease, scrofula

It is a chronic insidious contagious disease of man and animals caused by tubercle bacilli characterized by development of tubercles in lung tissue, lymph nodes or other organs resulting in caseation and calcification. The tubercles are frequently located in lungs and lymph nodes. But they may be formed in the liver, spleen, intestine, peritoneum, meninges and long bones. It is a zoonotic disease. Bovine tuberculosis, caused by Mycobacterium bovis (M. bovis) is considered as the serious epizootics of animals and humans. Bovine species are natural hosts to the disease, but a wide spectrum of domestic and wild animals, as well as man, can be infected. Animals reported as having been infected with M. bovis include North American bison (Bison bison), buffaloes (Syncerus caffer), elk (Cervus elaphus), domestic and wild pigs domestic (Sus scrofa), goats (Capra hircus), camels (Camelus bactrianus), dogs (Canis familiaris), cats (Fells catus), sheep (Ovis aries), possums (Trichosurus vulpecula), badgers (Meles meles), mink (Lutreola vison), ferrets (Putorius furo) and non-human primates. All species are not equally susceptible, and are often grouped into spill-over (end) hosts and maintenance hosts. Cattle and other bovine species are considered the primary and most well-known reservoirs or maintenance hosts. In countries where maintenance hosts are present endemically in the wild, infection from these populations to domestic cattle or other farm animals is difficult to avoid. In Africa, bovine TB primarily affects cattle; however, infection in other farm and domestic animals, such as sheep, goats, pigs, dogs and cats, is not uncommon. Wild ruminants and carnivores are also affected and are the natural reservoirs of the infectious agent in the wild. Man is also susceptible to the disease, the highest risk groups being individuals with concomitant HIV/AIDS infection. In Africa, human TB is widely known to be caused by M. tuberculosis; however, an unknown proportion of cases are due to M. bovis. This infection in humans is underreported as a result of the diagnostic limitations of many laboratories in distinguishing M. bovis from M. tuberculosis. TB is a neglected public health problem and accounts for about 25% of all avoidable adult deaths in developing countries. It is of socio-economic significance, as it can affect international trade in animals and animal products. Many factors account for the failure of developing countries to control and eradicate bovine TB, many of them politico-economic. Added to the high costs of a sustainable testing programme are problems of social unrest due to political instability and ethnic wars, resulting in the displacement of large numbers of human and animal populations; lack of veterinary expertise and communication networks; insufficient collaboration with bordering countries and hence a lack of quarantine; and smuggling of live animals across state boundaries. Scarce human and financial resources are often absorbed by action against the high incidence of other acute and fatal diseases, such as contagious bovine pleuropneumonia, foot and mouth disease, African and classical swine fever and parasitic diseases. The primary sources of infection for humans are consumption of unpasteurised milk and close association between humans and animals. Rural inhabitants and some urban dwellers in Africa still consume unpasteurised and soured milk potentially infected with M. bovis. Milk borne infection is the main cause of non-pulmonary TB in areas where bovine TB is common and uncontrolled.

Brucellosis, one of the most ancient diseases, continues to be a problem for humans and animals throughout the world causing enormous economic losses and significant human morbidity in endemic areas. The bacterial pathogen is classified by the CDC as a category B pathogen that has the potential for development as a bioweapon. It is one of the important zoonotic disease and considered as the most common laboratory acquired pathogen with >5 lakh human cases reported annually around the world. Brucellosis is a chronic infection which can result in infertility, delayed heat, interrupted lactation, loss of calves, wool, meat and milk production in animals and undulant fever in humans. Although in most of the counties, brucellosis is a nationally notifiable disease and reportable to the local health authority, it is underreported and official numbers constitute only a fraction of true incidence of the disease. Although the true incidence of human brucellosis is unknown, the estimated burden of the disease varies widely from <0.03 to >160 per 1 lakh population. Although only 17 countries in the world are free from the disease, it has been reported in many countries including India. Bovine brucellosis is endemic in all the states in India and appears to be on the increase in recent time, perhaps due to increased trade and rapid movement of livestock. The preponderance of natural bull service in rural India is perhaps yet another important factor in the maintenance and spread of infection. The Government of India has made it mandatory to regularly screen all the breeding bulls from artificial insemination centres, for brucellosis and to use brucellosis free bulls for semen production.

Etiology: Brucella sp. are facultative intracellular Gram - ve coccobacilli, nonspore forming and non-capsulated. It infects macrophages with principal involvement of liver, spleen and bone marrow. Nine Brucella species are currently recognized, seven of them that affect terrestrial animals are: B. abortus, B. melitensis, B. suis, B. ovis, B. canis, B. neotomae and B. microti and two that affect marine mammals are B. ceti and B. pinnipedialis. The first three species are called classical Brucella and within these species seven biovars are recognized for B. abortus, three for B. melitensis and five for B. suis. The remaining species have not been differentiated into biovars.

Synonyms: Calf scour, white diarrhea, septicaemia of calves

Colibacillosis is one of the important diseases in new born calves, piglets, lambs and foals caused by E. coli and characterized by marked postration, profuse diarrhea and septicaemia.

Etiology: Colibacillosis is associated with pathogenic serotypes of E. coli. Certain serotypes cause diarrhea and others cause septicaemia.

(a)  Enterotoxigenic E. coli: is the most common enteropathogen that causes diarrhea in new born farm animals. It causes bacteria by adhering to, colonizing and producing enterotoxins in the small intestine. They are not invasive. The enterotoxins produced by E. coli are of two types: heat stable (ST) and heat labile (LT).

(b)  Enteropathogenic E. coli: they attach tightly to the epithelial cells of the villus of small intestine. They do not produce toxins and seldom invade the intestine.

(c)  Enterohaemorrhagic E. coli: They are Shiga toxin or verotoxin producing strains of E. coli. They are not an important cause of diarrhea in farm animals but some produce diarrhea in calves. E. coli O157:H7, one of the important enterohaemorrhagic E. coli strains cause a broad range of clinical disease in humans including diarrhea in humans and haemorrhagic colitis and highly fatal haemolytic-uremic syndrome in children. 195

Glanders is a contagious, acute or chronic, usually fatal anthropo zoonotic disease of Equidae caused by Burkholderia mallei and characterized by serial development of ulcerating nodules that are most commonly found in the upper respiratory tract, lungs, and skin. The first recorded description of glanders is from the third century from Aristotle. In 1664, glanders was recognized as a contagious disease and in 1830, it's zoonotic potential was suspected. In the late 1800's, the Mallein test (1891) was developed for diagnosis of glanders. By 1910, the U.S., Canada and Great Britain all implemented glanders controlprograms. B. mallei was eradicated from the United States in 1929.

Etiology: It is caused by Burkholderia mallei. It is a straight or slightly bent Gm - non-spore forming, non-motile rod. The organisms in old culture appear as pleomorphic. The organisms can grow in ordinary media but addition of defribinated horse blood or glycerol accelerate the growth. Colonies have hop like odour and slime consistency. The organisms are readily destroyed by direct sunlight and most of the disinfectants. They can survive for 20 days in water and 6 weeks in contaminated stable. This organism is closely related to the agent of melioidosis, Burkholderia pseudomallei.

Susceptible hosts: The most susceptible hosts are horses, mules and donkeys. Infections can also occur in dogs, cats, goats and camels; cats may be particularly susceptible. Hamsters and guinea pigs can be infected in the laboratory. It is an occupational /professional hazard to human being. Laboratory samples are highly infectious to humans. Infection have been observed in sheep and goats. Guinea pigs, rabbits and field mice may be infected fatally. Cattle and pigs are absolutely resistant.

Geographic Distribution: Glanders is seen in some Middle Eastern countries, the Indian subcontinent, Southeast Asia, parts of China and Mongolia, and Africa. Sporadic cases are also seen in South America. Sporadic cases of glanders were identified between 1985-86 to 1990-91 from the states of Haryana, Himachal Pradesh and Punjab. Since reemergence of glanders in July - August 2006 in and around Pune and Panchgani area of Maharastra, several cases have been reported among equines of different states viz., Andhra Pradesh, Uttar Pradesh, Uttarakhand and Punjab. During the year 2006-07, National Research Centre on Equines, Hissar tested 4395 samples and 97 were detected positive and B. mallei was isolated from 8 of these cases.

Leptospirosis is a worldwide zoonosis. According to the occupational groups involved and the nature of the disease presentations, different names have been used, e.g. seven-day fever found commonly in Japan, Cane cutter's disease in Australia, Rice field Leptospirosis in Indonesia and Fort Bragg fever, which appeared as an outbreak in the US. Weil's disease, which is one of the severe forms of this disease, occurs in many countries, including India and other South-East Asian Countries, China, continental Europe and England. Leptospirosis exists in all the five inhabited continents and in a large number of countries. It occurs in tropical, subtropical and temperate zones. Leptospirosis is also an important cause of abortions, stillbirths and reproductive failures in cattle and pigs. Periodic ophthalmia in horse is commonly associated with leptospiral infections. Leptospirosis was first recognized in humans in 1886 by Weil and characterized by icterus. Leptospira have been isolated from a variety of animal species and humans manifesting various clinical syndromes such as mud fever and swine herd disease.

Etiology: The etiologic agent of leptospirosis is Leptospira interrogans. It is a thin spiral organism 0.1mm x 6 -20mm, with tightly set coils, and it is characterized by very active motility, by rotating ("spinning") and bending. Usually one or both ends of this single-cell organism are bent or hooked, but straight forms also occur that rotate and travel more slowly than hooked forms. The cells have pointed ends, either or both of which are usually bent into a distinctive hook. Two axial filaments (periplasmic flagella) with polar insertions are located in the periplasmic space. The structure of the flagellar proteins is complex. Leptospires exhibit two distinct forms of movement, translational and nontranslational. Morphologically, all leptospires are indistinguishable, but the morphology of individual isolates varies with subculture in vitro and can be restored by passage in hamsters. Leptospires have a typical double membrane structure in common with other spirochetes, in which the cytoplasmic membrane and peptidoglycan cell wall are closely associated and are overlain by an outer membrane. Leptospiral lipopolysaccharide has a composition similar to that of other Gram-negative bacteria, but has lower endotoxic activity. Leptospires may be stained using carbol fuchsin counterstain. Leptospires are obligate aerobes with an optimum growth temperature of 28 to 30°C. They produce both catalase and oxidase. They grow in simple media enriched with vitamins (vitamins B₂ and B₁₂ are growth factors), long-chain fatty acids, and ammonium salts. Long-chain fatty acids are utilized as the sole carbon source and are metabolized by b-oxidation. Because of their narrow diameter, the leptospires are best visualized by dark-field illumination or phase contrast microscopy and they do not stain readily with aniline dyes. The free living (L. biflexa) and parasitic leptospires (L. interrogans) are morphologically indistinguishable.

Synonyms: Circling disease, Meningoencephalitis, silage disease

It is an infectious fatal disease of wide range of animals and man characterized by encephalitis. It also causes abortion, endometritis and repeat breeding in farm animals. Historically, Listeria was first isolated in 1926 from a natural disease of rabbits characterized by mononuclear leucocytosis and therefore named as Bacillus monocytogenes by Murray and coworker at Cambridge. Later, Pierie who isolated similar bacillus from liver of gerbils in 1927 named it Listerella hepatolytica in honour of surgeon Joseph Lister and finally renamed it Listeria in 1940 due to taxonomic reasons.

Etiology: The disease is caused by L. monocytogenes. The genus Listeria consists of a group of Gm + bacteria of low G+C content closely related to Bacillus, Clostridium, Enterococcus, Streptococcus and Staphylococcus. Out of 6 species reported, only L. monocytogenes and L. ivanovii are considered virulent. Human cases of L. ivanovii infection are rare whereas L. monocytogenes is an important food borne pathogen. L. monocytogenes has been isolated from a variety of food products. Meat, poultry, dairy and vegetable products have been implicated as vehicles of listeriosis. The organism can be isolated from faeces and silage. The organism can be easily destroyed by ordinary disinfectants but may survive in soil and silage for a considerable period of time.

L. monocytogenes is small, coccoid with a tendency to occur in chains. It is aerobic or microaerophilic. It grows on ordinary media within a temperature range of 4-42°C. Growth is improved by addition of glucose, blood or liver extract. Colonies are β haemolytic on blood agar. It ferments glucose and salicin producing acid without gas. Lactose, maltose and sucrose are fermented slowly or not at all and mannitol is not fermented. It is catalase +. It shows tumbling motility when grown at 25°C but at 37°C is non-motile. It is because peritrichous flagella are produced by the bacillus optimally at 20-30°C but scantily or not at all at 37°C.