eChapters

38.1. Aim

To prepare nectar from different fruit pulp/juice.

38.2. Theory

Fruit nectar is also a RTS drink but it contains not less than 150Brix TSS and not less than 20% fruit juice or pulp. It is preserved only by heat processing as such no preservative is allowed in fruit nectars. Mango nectar, guava nectar etc. are the commercial products in this category.

38.3. Raw material, ingredients and utensils required

1. Mango, guava, and pineapple nectars are commercially available in the market. However the nectar can also be prepared from a number of other fruits like jamun, aonla, apricot, papaya, plum and bael etc.

37.1. Aim

To prepare ready to serve drink from fruits.

37.2. Theory

Ready to serve (RTS) drink is a type of fruit beverage which contains at least 10% of fruit juice and 10 degree Brix total soluble solids (TSS) with 0.3% acidity. It is not diluted before serving, hence it is known as ready to serve beverage. Mango drink, guava drink, pineapple drink are the commercial products available in the market.

Sweetened fruit juice is categorized as ready to serve beverage in which the minimum juice content shall be 85% with a minimum TSS of 10%, while unsweetened juice is a natural juice having 100% fruit juice with natural total soluble solids. These are preserved by heat processing method. Sweetened apple, pear juices are commercially manufactured.

36.1. Aim

To prepare cordial from lemon and lime.

36.2. Theory

Cordial is a sparkling, clear, sweetened fruit beverage from which pulp has been removed completely. It contains at least 25% fruit juice, 30% total soluble solids (TSS), 1.5% acid and 350 ppm sulphur dioxide. Lemon and lime juice cordials are the commercial products.

36.3. Raw material, ingredients and utensils required

1. Lemon and lime are used for making cordial.

35.1. Aim

To prepare squash from fruits or fruit pulp.

35.2. Theory

Squash is a type of fruit beverage containing at least 25% fruit juice or pulp and 40% total soluble solids (TSS). Squash is diluted before serving (1:3). According to FSSAI specifications, class II preservative in fruit squash shall not exceed 350 ppm sulphur dioxide/700 ppm KMS or 600 ppm sodium benzoate. Artificial colour and flavours can also be added when declared on the label.

35.3. Raw material, ingredients, machinery and equipments required

1. Fruits like mango, orange, lemon, lime, litchi, pear, apricot and pineapple are used for making squash.

34.1. Aim

To conduct cut out analysis of canned fruits and vegetable products.

34.2. Theory

Cut out examination of canned products is conducted to evaluate the product whether confirming to FSSAI standards or not. Canned fruit and vegetables are evaluated for presence of vacuum or pressure in the can, drained weight, total soluble solids, presence or absence of preservative, internal condition of can product contents and presence or absence of fermentation for ensuring microbiological safety.

33.1. Aim

To perform canning of fruit and vegetables.

33.2. Theory

Canning refers to a process which involves heating food stuff in hermetically sealed containers for a specific time at specific temperature to eliminate microbial pathogens that endanger public health as well as micro-organisms and enzymes that deteriorate food during storage. Broadly canning is a method of preservation of food for achieving longer shelf-life.

33.3. Principle of canning

Destruction of spoilage causing micro-organisms and enzymes within the sealed containers by means of heat.

32.1. Aim

To estimate the level of sulphur dioxide contents in fruit and vegetable products

32.2. Theory and principle

Sulphur dioxide in food product is added as class II preservative and may exists as undissociated sulphurous acid, as free bisulphate or sulphite ions or as combined SO2 in the form of hydroxy sulphonates. Sulphur dioxide is measured as free or total SO2. Estimation of SO2 in squashes, jam, RTS drinks is important as its presence beyond the prescribed FSSAI limits make the product not confirming to Indian Food Laws. Generally 2 parts of KMS (Potassium metabisulphite) releases one part of SO2 explained as under:

31.1. Aim

To estimate the sodium benzoate contents in fruit and vegetable products.

31.2. Theory and Principle

Sodium benzoate is used as a class II preservative in preservation of pickles, sauces, squashes etc. Its estimation is important as the quantity of class II preservative shall not exceed maximum limit prescribed under FSSAI. In a sample containing NaCl, the benzoic acid is converted into water soluble sodium benzoate by addition of NaOH. When the Sodium benzoate solution is acidified with excess of HCl, water insoluble benzoic acid is formed which is extracted with chloroform. The chloroform is removed by evaporation and the residue containing benzoic acid is dissolved in alcohol and then titrated with standard sodium hydroxide.

30.1. Aim

To estimate the salt contents in given samples by using Mohr method.

30.2. Theory and principle

Common salt is used during preparation of pickles, sauces, ketchup, fruit chutney and appetizers. Besides, imparting taste, the salt in appropriate concentration also acts as a preservative. In different products, it is estimated by using Mohr method. The sample is titrated with standard solution of silver nitrate for estimation of salt by using potassium chromate as an indicator. From the titre the concentration of salt is estimated by using a standard expression.

29.1. Aim

To estimate ascorbic acid in fresh and processed fruit and vegetable products.

29.2. Theory and Principle

Fruit, vegetables and their products are important sources of ascorbic acid. The ascorbic acid is present in sufficient quantity in aonla, guava, grapefruit, lemon, pineapple, strawberry fruits etc. Barbados cherry is the chief source of vitamin C. The products manufactured from these fruits are also considered as rich in ascorbic acid and the contents available in the commodities can be detected by using 2, 6 dichloro-phenol - indophenol visual titration method.

28.1. Aim

Estimation of reducing sugars in fresh fruits and processed products using Lane and Eynon method.

28.2. Theory

Sugars are inherently present in different fruits and vegetables or added during preparation of different products. They are added to the fruit products to improve the taste and also to act as a preservative. Glucose and fructose in the fruits represent reducing sugars while sucrose or cane sugar added represents the non-reducing sugar. They are estimated by using Lane and Eynon method which measures sugar as reducing sugar and total sugar as invert sugar.

27.1. Aim

To estimate the titratable acidity of fresh and processed fruit and vegetable products.

27.2. Theory

Most of the fruit, vegetables and their products contain acid or mixture of acids. The acids may occur naturally in the fruit and vegetables or may be added during manufacture of different products or by lactic acid or acetic acid fermentation. Generally citric acid is added in most fruit products while in pickles, sauces and ketchup acetic acid is used. The acids are mainly responsible for the tartness or sour taste, thus estimation of acidity is used as the measure of tartness. They also help in preservation by lowering the pH of the finished products.

26.1. Aim

To determine the pH of food samples.

26.2. Theory

The hydrogen-ion concentration of a food is a controlling factor in many chemicals and microbiological reactions. Pure water has equal concentration of H+ and OH- ions, each having the value of 1×107g/litre at room temperature that is why water is regarded as neutral. The pH of pure water is 7.0, the solution having pH below 7.0 is regarded as acidic while solution with pH above 7.0 is alkaline. Most fruits and fruit products have pH below 4.0 while vegetable; milk and meat products have pH more than 4.0. The pH value of some fruits and fruit products is given in Table 6.1. Thus, with the increase in acidity in the solution, the pH value decrease and vice versa. However, presence of buffer salts helps in maintenance of pH. Estimation of pH of the food is important as the processing conditions for different food products are categorized on the basis of their pH.

25.1. Aim

To estimate the ash content in fresh and processed products.

25.2. Theory and principle

Estimation of ash contents is important as it gives an idea about the presence of minerals in the food samples. Determination of ash is also required to evaluate fruit chutneys as the maximum limit for ash in this product shall not exceed 5% as per FPO specifications.

25.3. Apparatus/instruments/equipment

1. Crucibles

2. Muffle furnace

3. Sensitive balance

4. Heating plate

5. Desiccators

24.1 Aim

To determine the moisture content in fresh and processed products

24.2 Theory

Estimation of moisture in fresh and processed fruit and vegetable products is important for evaluating their nutritive composition as well as to optimize method for their processing like drying, preparation of different products viz; jams, preserve, candies etc. Moisture is determined by drying the weighed sample to a constant weight in hot air oven and measuring the loss in weight to express the moisture content in percent. This method is used for estimation of moisture in all food products which do not decompose at 100ºC. Moisture content of some commodities (Table 4.1) is as under:

23.1. Aim

To determine the total soluble solids in fresh and processed fruit and vegetable products.

23.2. Theory and principle

Total soluble solids (TSS) are determined by means of brix which measures the specific gravity or by using hand refractometer which measures refractive index. Brix is a measure of soluble solids only in case of pure sucrose solutions. Generally, the fruit juices contain more sugar than any other soluble constituent. Hence, brix provides a useful guide of soluble solids or sugar contents. Brix hydrometer is used to determine the percentage of brix in syrup while Abbe refractometer gives refractive index as well as degree brix. The Refractive index of the test solution is measured at 20°C using a refractometer and the soluble solid content is determined by the use of tables correlating refractive index with soluble solids. Generally, hand refractometer is used for determining TSS in food samples in fruit processing units (Table 3.1).

22.1. Aim

To estimate the fruit firmness.

22.2. Apparatus/machineries/equipments required

• Magness Taylor Pressure Tester

• Penetrometer

22.3. Theory and principle

Fruit firmness is an important feature of maturity. During ripening, fruits may soften and become too soft at the overripe stage. The changes are detected by touching or by using devices like texture analyzers and pressure testers to measure texture of fruits and vegetable.

21.1. Aim

To familiarize the students with the machineries and equipments used in fruit and vegetable processing.

21.2. Theory

The students will visit the processing unit/pilot plant and familiarize with different machineries and equipment used in fruit processing units. The students will observe the layout and the details of the machinery for different unit operations viz; raw material receiving, washing, sorting/grading, peeling, slicing, pulping, heating, filling, exhausting, sealing, sterilization/processing, labelling and storage for different products. The list of the machineries and equipments and important chemicals used in fruit preservation is as under:

21.3. Apparatus/machineries/equipments used in food industries

21.3.1. Basic equipment/ accessories

20.1. Introduction

Food laws and regulations are the legal standards of quality which regulate the manufacture and marketing of food products. The major objectives of enacting the food laws and regulations are:

1. To maintain quality of food produced

2. To prevent exploitation of the consumer by the sellers

3. To safeguard the health of the consumers and

4. To establish a standard of quality for different food products.

19.1. Introduction

Quality is a measure of the degree of excellence or degree of acceptability by the consumer. It is also defined as the combination of attributes or characteristics of the products that has significance in determining the degree of acceptability of the product to a user. Industry defines quality as the measure of purity, strength, flavour, colour, size, maturity, workmanship or any other distinctive attribute or characteristics of the product. According to USDA Marketing Workshop Report (1951) quality is the combination of attributes or characteristics of a product that have significance in determining the degree of acceptability of the product to the user . The quality standards of fresh and processed fruit or vegetable products vary with their intended use. For marketing purposes; size, attractiveness, maturity, organoleptic quality and freedom from defects are to be kept in mind. While for processing, physico chemical attributes of raw material such as presence of soluble solids, development of uniform colour, flavour, juiciness, uniform maturity, tenderness in some vegetables etc are taken into consideration. During processing of fruit or vegetables into value added products; colour, flavour and texture etc also become important.

18.1. Introduction

Spoiled food is defined as the food that has been damaged or has undergone changes so as to render it undesirable/unacceptable/unfit for human consumption. Spoilage of processed products is mainly caused by the action of micro-organisms and due to the physical or chemical changes. The reasons for microbial spoilage include under- processing, inadequate cooling, contamination from leakage through can seams and pre-process spoilage. In under-processed cans (receiving low heat treatment) large number of micro-organisms is expected to be involved in spoilage.

18.2. Causes of food spoilage

Food spoilage or deterioration may be described as loss of weight, softening, souring, rotting, wilting, moulding or a combination of one or more forms. The major causes of food deterioration are: biotic and abiotic factors.

17.1. Introduction

Tea (Camellia sinensis L), coffee (Coffea arabica L.) and rubber (Hevea brasiliensis) are known as estate crops or conventional plantation crops. These plantation crops are perennial in nature and cultivation of these crops generates considerable employment opportunities during on farm and off farm processing activities. The plantation crops are high value industrial crops contributing about 25% of AGDP. India is a largest consumer of tea accounts for 22% global tea consumption and also accounts for one-fifth of global tea area. Indian coffee industry also have prime place among plantation crops and also contributing 4% coffee to global market. Similarly, rubber plantations also provides basic raw material to toys, balloon, tyres, seed oil cake and rubber wood factory. The plantation crops along with main processed products also produce large quantity of biomass like husk, coir dust, coffee husk, tea waste, shedding etc which can effectively used to reduce nutrient requirements of crops. Parchment coffee prepared by using the wet method is generally favoured by the market. Cherry coffee, due to its very nature of preparation and longer contact with the mucilage and fruit skin, is usually associated with a characteristic fermenting flavour.

16.1. Introduction

Plantation crops are high value industrial crops grown in the tropics play a vital role in agricultural economy and export trade of developing countries. Plantation crops have great potential for utilization of waste land like rainfed dry land, hilly, arid and coastal areas to provide nutritional security, earn foreign exchange and to ensure livelihood security. The major plantation crops grown in India are tea (Camellia sinensis L), coffee (Coffea arabica L.), rubber (Hevea brasiliensis), coconut (Cocos nucifera), arecanut (Areca catechu L.), oilpalm (Elaeis guinensis), cashewnut (Anacardium occidentale L.), cocoa (Theobroma cacao L.) and cardamom (Elettaria cardamomum). India is the leading producer of most of these plantation crops. Among all fruit crops, the processing plays an important role in plantation crops as they are generally consumed only after processing. In addition to primary products which are exported largely, some by-products from these crops also have industrial as well as commercial uses. Coconut, arecanut, cashewnut, black pepper and cardamom are known as small holder plantations whereas, crops like rubber, tea and coffee are known as estate crops or conventional plantation crop. Coir, wood and shell from coconut, leaf sheath in arecanut, cashew nut shell liquid (CNSL) and feni from cashew are some of by-products with a great impact and demand in the market.

15.1. Introduction

Freezing is one of the oldest and most widely used methods of food preservation, which allows preservation of taste, texture, and nutritional quality in foods better than any other method. It is a process of bringing down the temperature of food below its freezing point and the frozen storage generally refers at temperature below -18oC.

15.2. Theory of freezing

The freezing process is a combination of the beneficial effects of low temperatures at which micro-organisms cannot grow, chemical reactions are reduced, and cellular metabolic reactions are delayed. Freezing is generally referred to as a unit operation in which the temperature of food is reduced below freezing point and a proportion of water undergoes a change in state to form ice-crystals. Immobilisation of water to ice and the resulting concentration of dissolved solutes in unfrozen water cause lowering of water activity of the food. Thus reduction in water activity and use of low temperature coupled with some pre- treatments like blanching is the basis for food preservation by freezing. Fruits such as strawberries, oranges, raspberries, black currants and vegetables like green pea, green beans, sweet corn, spinach, sprouts and potatoes are examples of commercially frozen products.

14.1. Introduction

Tomato is grown in India in abundance both in summer and winters. Tomato though botanically a fruit is generally considered as vegetable because of the way in which it is consumed. Fresh tomatoes are very refreshing and appetizing but can not be stored for a long period. During peak season, about 25% of the produce is spoiled due to mishandling and such losses can be avoided by converting tomatoes into different value added products. Tomato can be processed into pulp, paste, puree, juice, ketchup and sauce. In India, tomato sauce and ketchup are very popular. High quality products can be prepared from tomatoes by:

1. Using uniformly ripened, red coloured tomatoes.

2. Avoid prolonged heating and cool the product immediately after cooking.

3. Avoid using iron and copper equipments and utensils at any stage of processing.

13.1. Introduction

The preservation by using common salt, spices and vinegar is a common method for preparation of pickles, sauce and chutneys. Salt is mostly used as preservative in pickles in combination with acid. Minimum concentration of salt to act as preservative is about 12%. It inhibits enzymatic browning/discoloration by acting as an anti-oxidant. It exerts its preservative action by:

• Causing high osmotic pressure and thus suppress the microbial cells.

• Dehydrating food by tying up the moisture thus making it unavailable for the growth of microorganisms.

• Salt in the food affects the solubility of oxygen and thus growth of aerobic microorganisms is inhibited

• Chlorine in sodium chloride being good oxidizing agent is toxic to microorganisms

• Salt increases the permeability of the cell wall and thus cause changes in the permeability. Only the haloduric microorganisms can tolerate these effects while others are unable to survive.

• Similarly addition of acid to the food lowers the pH of the food which inhibits the growth of spoilage causing microorganisms.

• Addition of spices and edible oil in these products besides improving flavour and taste also help in preservation.

12.1. Introduction

Drying refers to the method of removal of moisture content from the food to a level at which the activities of food spoilage and food poisoning micro-organism are inhibited. Drying under open sun is probably the oldest method of food preservation used for agricultural crops including food grains, oilseeds as well as fruits and vegetables. Most of the fruits and vegetables contain enough moisture to permit the activity of enzymes and micro-organisms for spoilage and drying is necessary to reduce the water activity. Therefore, reduction in water activity of the food is the main principle of preservation by drying. Two commonly used terms for dried foods are low moisture foods and intermediate moisture foods (IMF). During drying, a single layer of fruit and vegetables, either whole or sliced after primary pretreatments is spread on trays which are placed inside the dehydrator or in the open sun for drying. In mechanical dehydrator, the initial temperature is generally kept at 43oC which is then gradually increased to 60-66oC for vegetables and 66-71oC for fruits.

11.1. Introduction

A fruit preserve is a product made from properly matured fruit by cooking it whole or in the form of large pieces in heavy sugar syrup, till it becomes tender and transparent. A fruit impregnated with cane sugar and glucose, followed by subsequent draining and drying is called a candied fruit e.g. petha candy, ginger candy. Further, candied fruit covered or coated with a thin transparent coating of sugar, which imparts a glossy appearance is referred to as a glaced fruit. When candied fruit is coated with sugar crystals either by rolling it in powdered sugar or by allowing the sugar crystals from glucose syrup to deposit on it, it becomes a crystallized fruit. The use of high concentration of sugar in these products acts as the preservative. The products like Petha candy, aonla preserve, apple rings; candied citrus peels, ginger candy, ginger in syrup; bael, pineapple and carrot preserve are the commercial products in this category.

10.1. Introduction

Sugar acts as a preservative by osmosis and sugar is not a true poison fo micro organisms. It absorbs most of the available water and restricts the availability of water/moisture required for growth of micro-organism. The concentration of 68 70% sugar is used for preparation of jam and jellies. Sugar adds value to these products as it imparts sweetness as well as body to these products. When sugar is boiled with acid and fruits it is hydrolyzed into dextrose and fructose, the degree of inversion depending on the pH and duration of boiling. Because of inversion of sucrose, a mixture of sucrose, glucose and fructose are found in jams and jellies.

Jams, jellies and marmalades constitute an important class of preserved fruit products. Apple jam, pineapple jam, strawberry jam and mixed fruit jam prepared by using pulp of two or more fruits are quite common. Similarly, guava jelly and orange marmalade are also examples of such products.

9.1. Introduction

Fruit juice and beverages generally comprise of naturally extracted juices, drinks, ready to serve (RTS) beverages, nectars, squashes, cordials and appetizers etc. These products are highly refreshing, thirst quenching, appetizing and nutritionally superior to many synthetic and aerated drinks. Fruit juice is the natural liquid expressed by pressure or other mechanical means from the edible portion of the fruit. Fruit juices are generally extracted from fruits in a number of ways, depending on their structure and composition.

The composition of juice is unaltered during preparation and preservation while for fruit beverages like drinks, squashes, cordial etc the fruit juice or pulp, sugar, acid, colour, flavour etc are mixed in appropriate proportions to a desirable taste. Apple juice, orange juice, mango squash, guava drink, pineapple juice and squash, mixed fruit drink, mango nectar etc are the commercial products available in the market.

8.1. Introduction

The term canning refers to a process which involves heating food stuff in hermetically sealed containers for a specific time at specific temperature to eliminate microbial pathogens that endanger public health and micro-organisms as well as enzymes that deteriorate food during storage. The canning of fruits and vegetables is canned out in the season when they are available in plenty. The canned products are then sold in the off season for getting better returns and to assure availability of fruits and vegetables throughout the year at reasonable prices.

8.2. History of canning

Important historical developments in canning are as under:

• The credit for invention of canning goes to Nicholas Appert, a French confectioner who was awarded a prize in 1809 by the French government for developing new method of heat preservation of food in sealed container and after whose name the process of canning is known as Appertization.

• In year 1810, Appert published the first book on canning entitled The Art of Preserving Animal and vegetable substances for many years which is the first known work on canning.

• The work of Appert consists of packing food in glass bottles, closing with corks and heating the container in boiling water for several minutes depending upon type of food.

7.1. Introduction

Preservation of fruit and vegetables by application of heat is one of the important methods of food preservation as it not only imparts desirable effect in the eating quality of the food but also exerts a preservative effect on the foods by destruction of micro-organisms, enzymes, insects and parasites. Other advantages of heat processing include destruction of anti nutritional factors (e.g. trypsin inhibitor in some legumes), improvement in availability of some nutrients (eg improved digestibility of proteins, gelatinization of starches, and release of bound niacin), production of shelf-stable foods that do not require refrigeration and above all it offers a relatively simple control of processing conditions. The disadvantages of heat processing include alteration or destruction of sensory quality of foods like flavor, colour, taste and texture as a result the heat processed foods are perceived to have different quality as compared to fresh produce.

6.1. Introduction

Chemical preservatives are those substances which minute organoleptic and physico-chemical properties . Preservation of food products with chemical preservatives is usually based on the combined or synergistic activity of several additives, intrinsic product parameters (e.g. composition, acidity, water activity) and extrinsic factors (e.g. processing and storage temperature).

Many chemical substances are added to the food for functional purposes and are called as additives and in many cases these are found to occur naturally in some foods. An additive is defined as a substance or mixture of substances other than basic food ingredient present in a food as a result of any aspect of production, processing or packaging.

5.1. Introduction

Food processing units range from home-based businesses to large-scale processing plants. The food processing sector is a highly fragmented industry which includes fruit, vegetables, dried, preserved, canned, pulps, pickles, chutneys, milk, dairy products, ready-to-serve products, frozen desserts, meat, poultry, marine products, grain processing, beer and alcoholic beverages, consumer, convenience and packaged food, soft drinks etc.

The main objective of fruit and vegetable processing is to supply wholesome, safe, nutritious and acceptable food to consumer throughout the year. Fruit and vegetable processing also aim to replace import of products such as squashes, jams, tomato sauces, pickles, etc., besides earning foreign exchange by exporting finished or semi-processed products. The fruit and vegetable processing industries are also established in the developing countries due to following reasons:

• Reduction in fruits and vegetables post harvest losses

• Industrialization policy of the government

• Generate both rural and urban employment

• Improve farmer s nutrition by self consumption of processed fruits and vegetables during off season

4.1. Introduction

Food occupies first position among the basic needs of human beings. Food processing dates back to the prehistoric age when crude processing including various types of cooking, such as over fire, smoking, steaming, fermenting, sun drying and preserving with salt were in practice. These crude processing techniques remained essentially the same until the advent of the industrial revolution. However, the change in food habits and the quality consciousness of the consumers in the developed world raises the need of advancements in development of food processing like spray drying, juice concentrates, freeze drying and also introduces dried instant soups, reconstituted fruit juices, and self cooking meals such as ready-to-eat food rations etc., during 20th century.

3.1. Introduction

Food preservation can be defined as the science which deals with the process of prevention of decay or spoilage of food thus allowing it to be stored in a safe condition for future use. Scientifically, preservation may be defined as a science which deals with the process for prevention of decay. All foods gradually undergo deterioration or spoilage from the time they are harvested or manufactured. Some commodities spoil rapidly, while others can keep for longer but limited periods. Spoilage in the food products may be caused due to some physical, chemical and microbial changes during storage. The preservation of these products can be resulted by controlling following physical, chemical and microbial changes.

2.1 Introduction

Food Pipeline refers to a post harvest system which deals with ensuring the delivery of a crop from the time and place of harvest to the time and place of consumption, with minimum loss, maximum efficiency and returns to all concerned including grower, processors and consumer. The term system represents a dynamic, complex aggregate of locally interconnected functions or operations within a particular sphere of activity. While, the term pipeline refers to the functional succession of various operations but tends to ignore their complex interactions.

2.2 Stages of postharvest system

The food commodities broadly undergo different stages/operations of post harvest systems right from harvest till consumption.

1. Harvesting

2. Threshing (food grains, pulses and oilseeds)

3. Drying (food grains, pulses and oilseeds)

Fruits and vegetables are important supplement to the nutritional requirements in the human diet as they provide the essential minerals, vitamins, flavonoids/ antioxidants and dietary fiber required for maintaining health. India is the second largest producer of fruits and vegetables in the world after China. During 2012-13, fruits and vegetables have registered a production of about 77.7 million MT and 159.5 million MT respectively (NHB, 2013. However, increase in production has not matched with the improvement in infrastructure for post-harvest management and processing. Fruits and vegetables are highly susceptible to deterioration owing to their high moisture content and delicate nature. Besides, most of the fruits and vegetables are seasonal in nature leading to glut in the local market during peak season resulting in huge economic losses to the growers. According to an estimate, 6-18% of fruits and vegetables are lost in the supply chain right from harvest to consumption due to lack of infrastructure for postharvest management, processing and value addition (Nanda et al., 2012).

Retrospection

India is a unique country in the world with many religions and cultural diversity. Indian culture has been continuing through the ages because of its adjustability with others and openness. Unity in diversity is one of the major characteristics of Indian culture. It is a diverse nation where people of many religions live together peacefully. Religion in India is characterized by the diversity of religious beliefs and practices. In Indian history, religion has played an important role in the cultural set up of the country.

Hinduism is the main Indian religion and considered as a universal order, a way of life which is followed by the people. The origin for Hinduism is very old having no exact date of its inception. This is a belief and practices followed by people through the ages and considered as a process of humanity. The earliest known sacred text of Hinduism is ‘Vedas’ which dates back to at least 3000 BCE. However, it is believed that these texts are of divine origin and timeless. Hinduism was more philosophically and theoretically described in Upanishads, written roughly between 800 and 400 BCE. The Mahabharata and Bhagavad-Gita are other best-known book and widely cited on Hinduism. The Ramayana is the other most important epic book on Hinduism which teaches about life, principles, duty, justice etc. through the life-story of Rama.

Dasharatha was a great Hindu king of the Kosala kingdom and Ayodhya was the capital from where He ruled. He was the son of King Aja of Kosala and Queen Indumati of Vidarbha. Dasharatha had three primary queens namely - Kausalya, Sumitra, and Kaikeyi. Unfortunately, the King had no sons to continue his dynasty after him. Despaired Dasharatha summoned Rishyashringa, a forest rishi for a ritual to fulfill his desire of have sons. By that way, the King had four sons. Queen Kaushalya gave birth Rama and similarly, Kaikeyi gave birth Bharata and Sumitra gave birth two sons, namely - Lakshmana and Shatrughna. Lakshmana was very much attached and devoted to Rama while Shatrughna was devoted to Bharata.

In previous Chapter, a list was furnished mentioning common names, botanical names, family and types of the all plants mentioned in the Ramayana. The purpose was to know the total number of plants and trees mentioned besides providing reference of the Shlokas as well. This information led to identification of the plants and trees but details about their morphological and floral characters were not mentioned. Therefore, in this Chapter details of the trees and other plants have been provided. This will help to know all about their botanical standing, conservation status, nativity, morphological and floral characters so that one can know the full details including present-day status, availability and importance (Cowen, 1950; Randhawa, 1983; Roy, 2015; 2024).

1. Botanical Name: Adenanthera pavonina L.

Synonym: Adenanthera gersenii Scheff.

Common Name: ‘Red Bead Tree’, ‘Curly Bean’, ‘Red Sandal Wood’

Family: Fabaceae

Nativity: India, Sri Lanka, Southeast Asia to Queensland

Conservation Status: Common (Not Threatened)

Type: Tree

Morphological and Floral Characters: Red Bead Tree is a timber tree found wild in India. Leaves compound, bipinnate, green when young, turning yellow on maturity. Flowers small, yellowish, grows in a dense, drooping almost cat tail like flower-heads. Fruits curved pods, hanging, green that turn brown, coil up and split open on ripening, showing small bright red seeds which are attractive, used as beads in jewellery.

In this chapter, a comprehensive list of all plants and trees mentioned in epic Ramayana has been furnished with salient details in order to establish their botanical identity and references of Sholka where it had been mentioned. Some of the plants and trees had been mentioned in multiple sholokas but the plant has been taken into account as a single reference to avoid repetition and to get correct count of the trees and plants in whole of the Epic (Amirthalingam, 2013, Arun Kumar, 2019; Balapure et al., 1987; Bast, 2022; Ghimire et.al, 2021; Mao and Dash, 2010; Prasad and Mishra, 1976).

The plant species mentioned in Ramayana were of different types having various usages. Considering their morphological characteristics, usages and other importance, they have been categorized broadly into the following heads.

1. Economically important plants (Timber value, product yielding etc. having commercial importance).

2. Edible (major and minor fruits etc.).

3. Culturally important plants (Plants used for plantation for shade, greening etc.).

4. Non-Edible Plants (Ornamental importance and other usages)

5. Medicinal Plants (having therapeutic uses).

6. Sacred Plants (having religious importance).

7. Others (having multipurpose uses).

The exile of Rama and his journey to the forests (vanvasa) for 14 years was started from Ayodhya (Uttar Pradesh, India) and ended to Lanka (Sri Lanka), through different forests, territorial regions, lands, hills, lakes of old India. The whole journey to those places was candidly portrayed in Ramayana with vivid descriptions, from narrations of the night sky, the position of different stars to descriptions of the flora, fauna and landscape. In this chapter, a retrospection of the places, forests, Rama passed through during his entire journey has been documented. The present status of those forests and places has been considered to evaluate a comparison over the thousands of years and the changes taken place besides significance they hold till date (Anonymous, 2024; Iyer, 2014).

Journey to the Forest and Route Map

Ayodhya, the capital city, kingdom of Kosala, was situated on the banks of holy river Saryu and was the birthplace of Rama. To fulfill the wish of his father, king Dasharatha, Rama left Ayodhya for 14 years of exile to the forest, along with Sita and Lakshmana. Their journey started from Ayodhya and passed through different places / forests.

The Ramayana written by sage Valmiki is one of the great epics of the Sanskrit language. It is most religiously regarded and popular book in Hinduism. The book is believed to be complied between 300 B.C.E. – 200 AD. However, the original story could be as old 500 B.C.E. (Goldman, 1984). There are many other versions of the epic in various regional languages found in India and South Asia (Raghunathan, 1981; Arya,1998; Dutt, 2004; Mukherjee, 2015; Sharma and Arya, 2022).

As per literature references, the name ‘Ramanaya’ is composed of two words, ‘Rama’ and ‘Aya?a’. Rama, the name of the main character of the Epic, has two contextual meanings. In the Atharvaveda, it means 'dark’, dark-coloured, ‘black' and is related to the word ‘ratri’ (night) which means 'darkness of night'. The other meaning, which can be found in the Mahabharata, is 'pleasing’, pleasant, charming, lovely, beautiful. The word ‘Ayana’ means travel or journey. Thus, Ramaya?a means ‘Rama's Journey’ from his kingdom to various places, forests during his life time (Anonymous, 1960; Tulsidas, 1966; Murthy, 2003; Kishore, 2007; Rajagopalachari, 2013).

Ramayana contains over 24,000 couplet verses, in its full form, which are divided into following seven ‘Ka??as’ (booklets) - Balaka??a, Ayodhyaka??a, Ara?yaka??a, Ki?kindhaka??a, Sundaraka??a, Yuddhaka??a, Uttaraka??a and about 500 sargas (chapters). It is regarded as one of the longest epic poems written in history (Keshavadas, 1988; Debroy, 2017; Gupta, 2021).

morally right man in the world? Narada replied that such a man is Rama, son of King Dasharatha of Ayodhya. Valmiki wanted to know more about Rama and proceeded to take bath when he saw two birds in playful mood and was happy. Suddenly, a hunter hiding in the bush shot one of the two birds. Valmiki became painful and chanted some dialogues in curse which ultimately created a new form of sloka. This was the beginning of the main narrative (Goldman, 1984; Sachithanantham, 2004).

India has a unique social and cultural setting with diverse people, culture and religions. It is one of the oldest and greatest civilizations of the world. Geographically India extends snow clad Himalayan mountains in the north to the tropical rain forests of the south. It is the 7th largest country in the world and well recognised in Asia by its mountains, seas, deserts, and vast stretch of fertile plain land, which give the country a distinct geographical identity.

India has a very rich, diverse religious and cultural heritage. The four main religions of India are - Hinduism, Islam, Christianity and Sikhism. Hinduism is the largest religion in India, with approximately 80% of the population is Hindu while Islam (Muslim) is the second-largest religion having 14% (approx.) share.

Religion in India is characterised by the diversity of religious beliefs and practices. In Indian history, religion has been an important part of the country's culture. Indian subcontinent is the birthplace of four of the world's major religions, namely, Buddhism, Hinduism, Jainism and Sikhism, which are collectively known as native Indian religions and represent 83% of the total population of India approximately.

Hinduism

Hinduism is the main Indian religion. It is considered as a religious and universal order, a way of life which followed by the people who believed in Hinduism. The word ‘Hindu’ refers group of particular people who believes and follow Hinduism, the oldest religion in the world. It has also been described as ‘Sanatana dharma’.

Epididymal Total Sperm Count

The epididymal sperm is obtained as described above, incubated at 35°C, which is the optimum temperature of rat epididymal sperm. The epididymal f luid is then diluted to a volume of 5 mL of pre-warmed (32°C) the spermatozoa are counted by hemocytometer using the Improved Neubaur (Deep 1/10mm. LABART, Germany) chamber as described by Pant and Srivastava (2003).

Total viable cell/ml = (Average number of sperm per chamber) x 103 x (Dilution Factor)

Liveability and abnormal sperm

Per cent live spermatozoa is estimated by differential staining technique using Eosin-Nigrosin stain (NE) (Campbell et al., 1953). These slides are also used for estimating the per cent abnormal sperm morphologically on the basis of observable abnormalities of head, neck, mid-piece and trail region of the spermatozoa.

The percent live spermatozoa are determined adopting the differential staining technique using Eosin-Nigrosin stain.

• A drop of epididymal semen is taken on a clean, grease free pre warmed glass slide.

• 4-5 drops of Eosin-Nigrosine stain (1% Eosin and 5% Nigrosine in 3% sodium citrate dehydrate solution) is placed near the semen drop.

• Epididymal semen and stain are mixed gently using a blunt fine glass rod.

• After 30 seconds to 1 minute a thin smear is made on a clean, grease free glass slide.

• The smear is examined under oil immersion objective.

Introduction

The immense use of chemicals and pharmacological entities has put the public health at brisk of potential health hazards. These substances in modern days are presented in the form of common constituents of food, medicines, beverages and junk foods etc. However, long term consumption of these substances might be responsible for chronic toxicity, but the accidental ingestion of large quantities of these substances may result into adverse acute toxic reactions. Hence an experimenter should have the acumen to develop and perform the acute toxic tests in order to find out the lethal dose 50 i.e. LD50 of the substances.

Acute Toxicity Studies

Acute toxicity studies helps to establish the dose dependent adverse effect including mortalities. The LD50 has been used as a major parameter in screening of pharmacological agents for acute toxic potential. Apart from mortality, other biological effects and the time of onset, duration and degree of recovery on survived animals, are also important in evaluation of acute toxicity. Hence, acute toxicity study solely gives information about LD50, therapeutic index and the degree of safety of a pharmacological agent.

Acute toxicity is defined as adverse effects occur following oral or dermal administration of a single dose of a substance, or multiple doses given within 24 hours, or an inhalation exposure of 4 hours. Oral administration is the most common form of acute systemic toxicity testing.

LD 50- It is defined as the concentration/dose that kills 50 % of the experimental population.

Acute toxic studies have been conducted in order to calculate LD50. There are several methods to calculate LD50, some are as follows:

Methods to Determine LD 50

• Karber’s method.

• Miller & Tainter method

• Lorke method

• Fixed dose method

• Acute Toxic Class method

• Up & down procedure

Karber’s Method

This method involves the administration of different doses of test substance to various groups, which has five animals each. The first group of animals is administered with the vehicle in which the test substance was dissolved or diluted

Isolation of Peripheral Blood Lymphocytes

Chemicals

1. Heparin

2. Sterile PBS (pH 7.2)

3. Histopaque-1077

4. RPMI-1640 Media

Protocol

Collect 5 ml of blood from the animal in 1 ml (20 IU/ml of blood) of diluted heparin and add 2 ml of PBS. Layer over the diluted blood on 5 ml of histopaque-1077 in 15 ml centrifuge tube and centrifuge at 1500 rpm for 40 min. Collect the buffy coat at the interface of plasma and histopaque in separate centrifuge tubes. Wash the cells thrice with PBS and suspended in 2 ml of RPMI-1640 media supplemented with 10% fetal calf serum. Determine the percentage cell viability by 0.1% trypan blue exclusion test, using hemocytometer. Finally dilute the cell suspension to 106 cells/ml in RPMI 1640 medium.

Pharmaceutical and chemical industries depend largely on toxicological evaluations to provide information regarding safety of their commercial products. Toxicologists so far have been relying on animal studies to investigate potentially beneficial or harmful effects of drugs and chemicals prior to testing them in humans. Traditionally, the safety assessment process during drug development has been based on screening indiscriminately a plethora of new chemical entities (NCEs) using a large number of animals as testing models in order to identify new therapeutic agents with the expectation that one or a few of these NCEs have some competitive advantages over the existing therapeutic agents. That is why they are more efficacious, bioavailable, selective, and safer. This approach is costly, time-consuming, and requires large quantities of test material. Currently, it takes about 10-12 years and a huge expenditure for a drug to move from research laboratory to patent. Thus alternative cell culture models in the early phases of the drug discovery process can save time and money both by eliminating least beneficial NCEs.

Preparation of Reagents

Culture media

For the culture of hepatocytes medium M199 (GibcoBRL, USA) is used. One pouch of the dry media is dissolved in 1 liter of filtered distil water. The pH of the media is adjusted by adding 0.4% sodium bicarbonate to be around 7.4. The media is than sterilized by filtration through seitz filter (0.22 mm) and stored in 50 ml sterilized conical flasks.

Acetylcholinesterase activity in brain is measured by the method described by Ellman’s et al. (1961).

Reagents

1. Phosphate buffer (0.1M) (pH 8.0) Solution A: 5.22 g of K2 HPO4 and 4.68 g of NaH2 PO4 are dissolved in 150 ml of distilled water. Solution B: 6.2 g of NaOH is dissolved in 150 ml of distilled water. Solution B is added to solution A to get desire pH 8.0 and then finally the volume is made up to 300 ml with distilled water.

2. DTNB reagent 39.6 mg of DTNB with 15 mg of NaHCO3 is dissolved in 10 ml of 0.1 M phosphate buffer.

3. Acetylthiocholine (ATC) 21.67 mg of Acetylthiocholine is dissolved in 1 ml of distilled water.

Procedure

The brain tissue is weighed and 10% homogenate is made in 0.1M Phosphate buffer (pH 8.0). Take 0.4 ml aliquot of homogenate in a cuvette containing 2.6 ml phosphate buffer (0.1M) and 100 µl DTNB. The contents of the cuvette are mixed thoroughly by bubbling air and absorbance is measured at 412 nm in a spectrophotometer. When absorbance reached a stable value, it is recorded as the basal reading .To this 20 µl of acetylthiocholine is added and change in absorbance is recorded for a period of 10 min at intervals of 2 min. Change in the absorbance per minute determined.