eChapters

Introduction

Tissue culture refers to the technique of growing cells, tissues, or organs under controlled, artificial conditions. This allows propagation of tissues from a small sample of cells/tissues under sterile conditions. Plant tissue culture is widely used for various purposes, including the production of disease-free plants, rapid multiplication of plants with desirable traits, genetic modification, conservation of rare or endangered species, etc. Tissue culture is a valuable technique because it enables consistent and large-scale production of plants, ensures genetic uniformity, and allows manipulation of plant biology at cellular level. In this chapter, we will discuss about the major types of tissue culture techniques and their advantages, disadvantages, as well as applications in brief.

Different Types of Tissue Culture

Callus culture involves induction and growing of unorganized, undifferentiated cell masses from an explant on a nutrient/culture medium. The part of plant used to induce callus is called explant which can be tissue, organ or part of a plant.

Organ culture is used to maintain or grow an entire organ or a part of organ in vitro. It preserves the structure and function of the organ for studies or transplantation.

Embryo culture involves isolating and growing embryos from seeds or ovules in a nutrient medium. It is used to bypass seed dormancy or rescue embryos from hybrid crosses that may not develop naturally.

Cell suspension culture involves growing dispersed cells or small cell clusters in a liquid medium under continuous shaking. This is used for large-scale production of cells, secondary metabolites, and studying cell biology..

Introduction

Different types of culture media are used for culturing microbes and plant tissues. Preparing as well as sterilizing them properly following appropriate method with safety for the desired purpose are important parts of research work. In this chapter, we will discuss briefly about different types of media, their composition, preparation and sterilization in laboratory for microbial or plant tissue culture.

Culture Medium

Ingredients mixed to prepare solid, liquid or semi-solid substance to support the growth of microorganism or plant tissue in vitro (outside the body) is called culture medium. Frequently, it becomes important to grow microorganism or plant/animal tissue in vitro for any or all of the following purposes:

• To study characteristic features or properties of microorganisms.

• To identify the causal organism(s) of infection in a clinical sample, so that appropriate remedy can be determined.

• To culture/multiply microorganism or plant/animal tissues in vitro for research or commercial purposes.

• To prepare/produce biological molecules, vaccines, antigens, proteins, metabolites, etc. in vitro for research or industrial purposes.

Introduction

Use of acid/base is common in laboratories for experiments like titration and proper safety in their handling laboratory is necessary. In this chapter we will briefly discuss about the safety measures necessary to be adopted while using acids/bases in laboratory for any experiment.

Handling of Strong Acid/Base

There are four major points that need to be taken care of while working with strong acids or bases that might be quite harmful if any accident happens.

1. Protect Yourself

Wear a lab-coat and acid-resistant gloves: Always use lab-coat/apron and make it sure that the sleeves properly cover your arms. More importantly, use gloves made of acid-resistant materials and also wear shoes to protect your feet while wearing long-pants/jeans. Female with long hair, must tie her hair properly (with rubber band) to avoid any disturbance caused by appearance in between.

Wear safety goggles: It is important to wear safety goggles to protect your eyes using larger and fitting goggles that cover front as well as sides of your eyes. Always use properly fitting and adjustable goggles because smaller/ loose goggles may not protect the areas around your eyes against accidental acid spills and it may fall down from your face, if it is too loose.

Locate emergency wash area: There must be an emergency washing area nearby the work place or laboratory containing lab spill management kits. All the workers in the laboratory must be aware of using the kits and emergency plans.

Neutralize acid: Don’t panic even if an acid or chemical spill happens. Stay calm and act as quickly as possible. In case an acid spills, neutralize it by using sodium bicarbonate so that they can be cleaned and disposed safely. If acid comes in contact of skin, wash it off under running tap water but do not use/ apply sodium bicarbonate on skin.

Introduction

Measurement of pH of a solution and preparation of different types/strength of buffers are common practices in a laboratory, particularly in the Biochemistry and Molecular Biology laboratories. Therefore, in this chapter you will be acquainted with the procedures of measuring pH of a solution as well as preparation of some of the commonly used buffers in a laboratory.

Potential or Power of Hydrogen Ion (pH)

Measurement of pH of a solution is measuring the molar concentration of hydrogen ion [H+] in the solution, which is an indication of the acidity or basicity of the solution. The symbol pH stands for “power of hydrogen” and numerical value for pH is just the negative logarithm of the molar concentration of hydrogen ions [H+]. The ionic product of water (Kw ) is the basis for the pH scale. The total hydrogen ion concentration from all the sources and it can be experimentally measured.

The pH scale designates the concentration of H+ (and thus of OH−) in an aqueous solution, which ranges between 1.0 M H+ and 1.0 M OH−. The term pH is defined by the expression

Thus, the ionic product of water makes it possible to calculate the concentration of H+, with given concentration of OH−, and vice versa. The value of 7 for pH of a neutral solution is not an arbitrarily chosen figure; it is derived from the absolute value of the ionic product of water. A solution having pH greater than 7 is basic, and the concentration of OH− is greater than that of H+. A solution having a pH less than 7 is acidic.

Introduction

For agricultural applications as well as for research work, often we need to prepare solutions of different agrochemicals of varying strength, as per the requirements. Agrochemicals are used to help crop plants grow, produce, and protect them from biotic/abiotic stresses. Many of the agrochemicals (insecticides, herbicides, fungicides, rodenticides, molluscicides, nematicides, etc.) are used to protect the crop from insect pests and diseases. However, some of them (fertilizers and soil conditioners) are used to augment crop yield. While in the previous chapter (Chapter 8) we discussed about preparation of laboratory solutions, in this chapter we will briefly discuss about the preparation of different agrochemicals.

Agrochemical/agrichemical is a generalized term used for the chemicals applied in agriculture. In general, it also includes the feed-supplements used in f isheries and animal husbandry.

Agrochemicals

Fertilizers

Fertilizers are used to augment growth of crop plants and to mitigate nutrient deficiency in the soil. Fertilisers can be organic (containing natural substances, prepared through natural processes) and inorganic fertilisers (synthetic fertilizers, manufactured artificially using chemical processes using natural deposits, by chemical alterations).

Soil Conditioners

Soil conditioners include manure, livestock and leaf compost, peat that help making the soil in good condition. They are laid on the top soil and then mixed properly. They not only increase nutrient contents in the soil but also improve aeration and water holding capacity.

Introduction

In a research laboratory, we frequently need weigh chemicals and prepare different solutions of varying strength, as per the requirements. Accuracy in weighing and preparation of solution, with respect to weight/volume and strength, determines the success of experiment. In turn, the accuracy depends on weighing of solid chemical and/or measuring the volume of liquid chemical/making up the volume. In this Chapter, weighing of chemicals using appropriate weighing balance, preparation of solutions of different strength, their dilution, and precautions while preparing will be discuss in brief.

Weighing of Chemical

The primary method to determine the amount of chemicals to be used is weighing the chemical by using a balance.

In general, there are two types of balances to weigh chemicals in a laboratory:

1. Standard laboratory balance

2. Analytical balance

Standard Laboratory Balance

Generally, a standard laboratory balance is used for weighing a chemical when an error of ±0.02 g is acceptable. It is also used for pre-weighing, where approximate mass needs to be determined. Precise mass of the chemical is then determined using an analytical balance. Following are the steps in using a weighing balance in a laboratory.

Introduction

To accomplish experimentations in laboratories, sometimes we need microscope to test/validate our ideas/findings. A microscope is needed to examine objects which are very small which cannot be visualised by naked eyes. However, microscopic study requires some expertise in operating the microscope, particularly the sophisticated microscopes, as well as to prepare (peeling, sections, cutting, fixation, staining, dehydration, mounting, etc.) the sample, so that it can be appropriately examined for the anticipated details.

According to the scientific requirements different types of microscopes are used in the laboratories for research purpose. In this chapter, our aim is to acquaint you with the basics of using and handling of microscopes, particularly a compound microscope that is commonly used by the graduate and post graduate students for basic research.

Microscopy is often designated as the scientific investigation that is employed to review and analyse minute structures and objects using a microscope. The term “microscope” has been derived from ancient Greek words “micros” meaning “small” and “skopeîn” meaning “to look at or see,”.

Microscope

A microscope is a laboratory instrument used to examine objects (biological specimens) which are too small to be seen by the naked eye. The microscope must accomplish three tasks:

• Magnification- to provide a magnified image of the specimen.

• Resolution- to separate the details in the image.

• Contrast- to render the details clearly visible to the eyes.

Although we will tnroduce to you the different types of microscopes used for different speciments, our main focus is to discuss about compound microscope, which is commonly used in the laboratories. Compound microscope was developed around the beginning of 17th century.

Since mouth pipetting of liquid reagents in a laboratory is not advisable, different tools are needed for safe, easier, and precise dispensing of liquid reagents. This chapter is devoted to make you aware of the tools like vacupette and micropipette which can be used for the handling of liquid reagents in a safe and precise way.

Vacupette

Glass pipettes have been traditionally used by sucking liquid solvent/solution directly using mouth. For safety reasons, mouth pipetting is no longer permitted in a laboratory. Hence, to fill a pipette vacupette is used by placing it on the top of glass or plastic pipette. It acts as a device to create vacuum for filling reagent in the pipette as well as for releasing the reagent with mechanical regulation on the flow out of the liquid. Vacupette can be of different types.

1. Rubber Bulb Vacupette

Rubber bulb vacupette is often used to transfer (pump up and discharge) liquid using a pipette when a larger quantity of chemicals is to be transferred. Small/tiny rubber bulb is attached to glass/plastic pipette for drawing little amount of reagents. Small vacupette is used to precisely discharge smaller volume/ drops.

Handling of Vacupette

• Insert the top of a pipette in the bottom of the rubber bulb vacupette (Figure 6.1).

• Remove air from the pipette by squeezing valve “A” at the top of the vacupette while simultaneously squeezing the rubber bulb. The amount of air released depends on the size of the rubber bulb.

Different basic as well as advanced laboratory equipment need to be used in a laboratory while conducting experiments. Many of them like pH meter, laminar air-flow hood, viscometer, thermometer, magnetic stirrer, microwave ovens, incubators, sand-bath, water-bath, oil-bath, etc. are the basic and simple equipment which require to be used routinely while conducting experiments in a laboratory. However, basic knowledge of their operation, handling procedure, when to be used, how to be used, and more importantly what not to be done with it, etc. are essential for keeping them in serviceable condition as well as for the accuracy/precision in their operation. In this chapter, we will discuss about some of the routinely used laboratory equipment along, while some other sophisticated equipment will be discussed in other chapters.

pH Meter

A pH meter measures exactly the H+ concentration of a solution; thus indicates acidity or alkalinity of a solution with greater precision. Actually, it measures the difference in electrical potential between a pH electrode and a reference electrode, and display the results converted into the corresponding pH value. Electrode is the key part (rod-like structure, usually made of glass) of a pH meter, having a bulb at the bottom containing a sensor. The pH meters range from simple and inexpensive pen-like devices (portable/pocket pH meter) to complex and expensive laboratory instruments with computer interface (Figure 5.1).

Frequent calibration of a pH meter using the solutions of known pH (standard solution of pH 4.01, 7.00, 10.01, Figure 5.2), generally before each use, ensures the accuracy of the instrument. For very precise work the pH meter should be calibrated before each measurement. For normal use calibration should be performed at the beginning of each day. This is necessary because the glass electrode does not give a reproducible measurement over a longer period of time.

A variety of glassware like burettes, pipettes, measuring cylinders, flasks, separating funnel etc. are used for experimental purposes in a laboratory. All of these need to be used and handled correctly, otherwise the chances of experimental error and damage to the glassware increase significantly. Being made of glass, they need special care while washing and using in experiments because even a minor mistake in handling of glassware may result in breakage of the apparatus. With this precautionary note, let us discuss about some of the specific glassware used in the laboratories.

Burette

Burette is a long graduated glass tube with a stopcock at one end for delivering a known/desired volume of a liquid, especially in titration experiments. It contains a stopcock at the lower end and a tapered capillary tube as the stopcock’s outlet. The flow rate of liquid from the tub stopcock’s outlet can be controlled with the help of stopcock valve. Burette is used for measuring the volume of a solution/chemical added in a quantitative (titrimetric) estimation experiment.

Procedure of using a Burette

The burette reading must be noted before and after delivering the chemical/ solution from the burette. The difference in these two readings is the volume of the chemical/solution released out of the burette. The solution should be released slowly/drop-by-drop to avoid any experimental error/mistake. If the solution/chemical is allowed to pour out too fast, the wall of the burette will not drain properly and some liquid may remain sticking to the surface of the walls. Moreover, excessive amount of chemical may be released/dropped during the experiment, particularly during the titration experiment wherein we need to know the exact amount of acid/base needed to get neutralization point. Thus, any minor mistake may lead to get a faulty reading. The measuring capacity of a burette usually used in a laboratory is 50 mL.

Introduction

A variety of glassware, plasticware, and other apparatus are used for experimental purposes in a laboratory. They need to be washed, cleaned, dried, and sometimes sterilized before they can be used in the next set of experiments. Glassware needs special care of handling during washing, drying, sterilization, etc. because even a minor mistake in handling of glassware may result in its cracking/breakage and it may become unserviceable. All of these may result in a delay in experimentation if spare piece of the glassware is not available or it may affect serviceability of the associated apparatus.

A large number of glassware such as beaker, flask, measuring cylinder, burette, test tube, etc. are regularly used in a laboratory, which requires careful handling during the different stages of experiment as well as after completion of the experiment while preparing them for another set of experimentation.

Glass being a complex silicate, its properties depends on the type of silicate anions in the structure and the cations formed. Thermal properties of glass changes by the addition of boron oxide (B2 O3 ); hence, borosilicate glass is extensively used in the laboratories. With this brief background information, let us discuss about the necessity, procedures, and precautions during washing, cleaning, drying and sterilization of laboratory-wares. Figure 3.1 shows various advantages of using borosilicate glass in a laboratory.

Good laboratory practices require clean glassware, because an experiment may give erroneous result if dirty glassware is used at any step of the experiment. Hence, the glassware must be physically and chemically clean; and in many cases, it must be microbiologically clean or sterile. The commonly used glassware in a laboratory include test tube, measuring cylinder, pipettes, beakers, flasks, microscopic slides, etc.

Introduction

While experimenting with chemicals, sometimes we need to dry the chemical/ solution (solid, liquid or gas) using a process like evaporation or distillation, mainly to remove water or different solvents. Drying is the process of evaporation or distillation to remove water/solvent from a solution, solid or even from gas. Generally, a drying agent (usually an anhydrous inorganic salt) is used for this process, which reacts with the water (present in the chemical/ solvent to be dried) to form a hydrate. For example, anhydrous MgSO4 is one of the drying agents. The drying agents are recognized by their capacity (the rate at which they absorb water) and their intensity (the amount of water left behind in the solvent at equilibrium). No chemical reaction or change in phase occurs during the drying process.

Type of Solvents

Apart from water (a universal solvent), there are three families of organic solvents used in laboratories.

1) Oxygenated solvents: Alcohols, Ketones, Glycol ethers, Glycol esters, etc.

2) Hydrocarbon solvents: Aliphatic and Aromatic hydrocarbons.

3) Halogenated solvents: Chlorinated hydrocarbons.

Introduction

For proper and efficient experiment to be conducted, experimenters need so many chemicals to deal with. This includes different acids, base, salt, different kind of indicators and so on. Some of these chemicals are safe for health, but some of them are dangerous to health when come in close proximity to our body while some of the chemicals cause health issues on inhaling. These may cause improper functioning of some of our physiological activities. Acids and bases are very dangerous for our tissues when come in our contact. These may cause serious respiratory problem in addition to injuries on coming to our contact. Some chemicals are exploratory yet they may cause even death of the person working with them in laboratory. Therefore, mere use of the chemicals in laboratory is not only the objective of the experimenter. The person working in laboratory should also be concerned about the proper use and handing of the chemical while working with it. Thus, every researcher of an institution who has to work with chemicals in a laboratory should be made aware about the chemicals to be used in the laboratory and the consequences if any mishandling of a laboratory chemical occurs. Not only these, suitable precautionary measures should also be known to the researcher along with ensuring the availability of the required safety equipment/installations. Hence, a proper understanding of the ways of handling of chemicals in a laboratory is of prime importance for the personals like you who are working or have to work in laboratory.

Safe Handling of Chemicals in Laboratory

Safe use/handling of chemicals in laboratory include minimized exposure to chemicals, maintaining safety data sheets, proper labelling, storage, segregation, and transport of chemicals for personal hygiene.

1. Calibration of Milk Butyrometer

Introduction

Butyrometer is used for determination of fat for milk and milk products by Gerber method. The milk butyrometer is a graduated vessel, which are designed in such a way that the internal volume of each unit is 0.125 ml which is corresponding to 1 percent fat. Sometimes due to manufacturer’s error, the volume of one unit of milk butyrometer is not exactly equal to 0.125 which leads to subsequent error in results in milk fat. So before starting to use a new butyrometer, it is essential to calibrate for its efficiency in the laboratory. Mercury is used to calibrate milk butyrometer.

Apparatus

Milk butyrometer (0 – 10% graduation), analytical balance

1. Preparation of Standard Sodium Hydroxide Solution

 Introduction

 When oxalic acid reacts sodium hydroxide, sodium oxalate and water are

form. Phenolphthalein indicator detects the end point by its characteristics

colour change.

 Apparatus

 Burettes, pipette, volumetric flask, beaker

 Reagents

 • Oxalic acid crystals

 • Sodium hydroxide – 0.1 N approximate

 • Phenolphthalein solution – 0.1 percent in ethyl alcohol

Procedure Preparation of 0.1 N oxalic acid solution 1. Weight accurately 1.575 g of oxalic acid into a 250 ml beaker using a weighing bottle. 2. Add some distilled water to the beaker and dissolve the oxalic acid with the help of glass rod. 3. Transfer the solution into a 250 ml volumetric flask through a funnel. 4. Wash the beaker 3- 4 times with small quantities of distilled water and transfer the washing into the flask. 5. Make up the volume up to the mark with distilled water and mix thoroughly. 6. Preparation of approximately 0.1 N sodium hydroxide solution 7. Weigh about 4.25 g of sodium hydroxide pellets and dissolve into 200 ml of distilled water.

1 Detection of Starch Introduction

Starch is widely available in several forms such as wheat flour, corn flour and commercially manufactured starch. It is extensively used by adulterators to raise the SNF content in the milk. When high amounts of starch are added to milk, it can cause diarrhea due to the indigestion of starch in colon. Apart from this, accumulation of starch in the body may prove very fatal for diabetic patients. In this method, starch in milk can be detected by using iodine solution. Iodine from the iodine solution gets absorbed on the starch molecule surface and resultant product of adsorption has deep blue colour.

Reagents

 • Potassium iodide

 • Iodine solution - 1% (Add 1 g of iodine and 5 g of potassium iodide in

distilled water and dilute to 100 ml)

Sample Preparation

Sample should be taken at six or more points on the top of the surface by means of a tubular trier of sufficient length to extend to the bottom of the barrel. Remove this cores and transfer to clean, dry, air tight container and seal immediately.

Before opening sample for analysis, make homogenous either by shaking or by alternately rolling and inverting container. If lump are present, sift sample through No 20 sieve, rubbing material through sieve and tapping vigorously, if necessary.

Avoid sampling on rainy day or when humidity is high, so as to reduce moisture absorption from air.

1 Determination of Moisture Content by Moisture Balance Introduction

In this procedure, infra-red radiation is used to accomplish rapid drying. The tester has a sensitive torsion balance which permits rapid determination directly from the balance scale, eliminating the need for computing or calculation to obtain results. This method is useful for the in-process determination of moisture because the results may be obtained in minutes.

Sample Preparation

a) Plain products: Sample should be softened at room temperature. Since the melted fat tends to separate and rise to the surface, it is not advisable to soften the sample by heating on a water bath or oven or flame. Mix thoroughly by stirring with spoon or eggbeater or by pouring back and forth between beakers.

b) Ice-cream containing Fruits and Nuts: Fill one third part of the cup with ice cream sample. Melt the product at room temperature and mix until insoluble particles are finely divided (about 3-5 minutes for fruit pieces and upto 7 minutes for nut pieces). Otherwise, the product may be ground in a porcelain or glass pestle and mortar. Don’t let temperature exceed 25oC at any time during softening and mixing steps. If fat separation occurs, discard sample otherwise immediately pour mixture into wide jar and cap tightly. If allowed to stand, shake vigorously before taking sample for analysis.

Sample Preparation

 Cheese sample is prepared by passing through 8 mesh sieve three times or

grate sample and mix thoroughly or grind to a uniform mass in a glass pestle

and mortar. Keep the sample in an airtight container until the time of analysis.

 1 Determination of Moisture Content in Cheese

 Introduction

The moisture content of cheese is defined as the loss in mass, expressed as a

percentage by mass when the product is heated in a hot air oven at 102 ± 2°C

to constant mass.

1 Analysis of Cream

 Sample Preparation

Warm the cream sample to 40 - 50oC if the sample is very viscous. Then pour from one vessel to another 3 to 4 times to mix well. Sample should be tested daily and always within three days after collection since enzymatic deterioration of the cream will cause low tests.

1.1 Determination of Fat Content in Cream

 1.1.1 Gerber Method

 Introduction

Sulphuric acid is added to cream in order to dissolve the protein and other solids and create heat which liquefy and de-emulsify the fat. Iso-amyl alcohol is added to aid the separation of fat. Upon centrifugation, fat being lighter will be separated on top of solution in the cream butyrometer.

1 Analysis of Chemical Composition of Milk

 1.1 Determination of Acidity of Milk

 Introduction

The acidity of milk can be determined by titrating the milk against standard sodium hydroxide solution using an indicator like phenolphthalein and is expressed in the terms of lactic acid. Though natural acidity in milk is due to its constituents such as casein, albumin, citrates, phosphates and carbon dioxide but it is customary to express in terms of lactic acid. As one gram equivalent weight of an alkali neutralizes one gram equivalent weight of an acid, the volume of standard alkali required to neutralize lactic acid of fixed volume of milk gives percent acidity of milk according to the reaction.

Milk, commonly referred to as the “ideal food,” is an essential source of nutrition for human consumption. Because of the growing human population and the rising demand for milk and milk products, there is more interest in and concern about the quality and safety of dairy products. Milk comprises chemical substances with various functions in the human body, such as carbohydrates, lipids, proteins, and trace elements (vitamins, minerals, etc.) (Guetouache et al., 2014).

The exploration of facts about healthy and nutritious food has increased over the years. The advent of advance scientific tools has elucidated facts regarding the role of diet or nutrients in regulating gene expression (Sales et al., 2014). Various studies based on molecular interactions in food has revealed that many dietary components such as proteins, carbohydrates, fats, minerals, vitamins and phyto-constituents has the tendency to regulate gene expression.

The term nanoscience defines controlling or modifying the shapes and sizes of the materials such that they lie in the range of a nanometre (1 nm = 10^-9 m) scale, typically 1-100 nm (Chavada et al., 2016). The application of these materials and systems by their suitable designing and production is what is known as nanotechnology. Richard Feynman, in 1959, first discussed the possibility of this advanced technology. However, the term was first used by Norio Taniguchi in 1974 (Chavada et al., 2016).

Starch is the most abundant polysaccharide carbohydrate reserve which is available in different parts of plants such as roots, stem, leaves, seeds, flowers, and fruits. It is considered an excellent source of energy and plants utilizes carbon from it for their metabolic activities (Smith, 2001). It is composed of glucose units linked together by glycosidic bonds to form amylose and amylopectin polymeric chains (Abbas et al., 2010).

Enzymes (biological catalyst) can be defined as a substance which initiates or fastens the reaction rate, without being consumed during the reaction. Enzymes function as a catalyst in various biological processes taking place in all life forms. Although enzyme synthesis takes place inside living cells, they can even function as a catalyst in the in vitro processes, which makes it suitable for many industrial applications. For product formation in any chemical reaction, reactants or substrates has to overcome the transition state.

Fourier transform infrared (FTIR) spectroscopy is one of the most effective methods for determining the functional groups and potential molecular interactions between membrane chemical molecules. Understanding of (infrared) IR absorption band placements in the spectrum as wave numbers can be used to identify the various chemical constituents (such as aromatic amides) that may not be visible by X-ray photoelectron spectroscopy.

Biosensors are the next generation detection devices, use biological sensing elements like enzymes, nucleic acids, antibody, cells, organelles, tissues, receptors; have got varied applications in the field of drug discovery, diagnosis, biomedicine, food safety and processing, environmental monitoring, defense and security etc. Microbial biosensors are analytical device that uses naturally occurring or engineered microbial cells as the recognition elements to detect physiological changes in cells with the help of suitable physicochemical transducer.

The last two decades have seen rapid growth and development in the industrial sector as well as urbanization has increased. Due to globalization and mass production of commodities, the situation in the modern world nothing like ever before. Food and dairy industries are also not left out from this massive expansion. Due to changing behavior and food consumption habits too, there has been a surge in the risk of foodborne illness. This has created a problem in terms of public health and sustainability

Thermal heating methods are used from long time to decontaminate the food products from microorganism and therefore increase its shelf life and ultimately make the product safe for human consumption. But these methods diminish the sensory, physicochemical and nutritional properties of food (Barba et al., 2012). Also, the cost of generation of heat in manufacture of food product make the product costlier in thermal processing and also create environmental issue.

Electricity is one of the fundamental phenomena of nature. Electricity or electric current is caused due to the movement of charged particles i.e., electrons. Due to the potential difference between two points in a conductor, electron movement takes place. The potential difference between two points is caused due to the difference in charge density. Electron tends to flow from lower potential to higher potential which causes electricity to flow. In general, the electrons flow from cathode (negative) to anode (positive) but in a wire electricity, electrons flow from anode to cathode when connected to a battery or any electricity source.

High-pressure processing (HPP) is a novel method for non-thermal preservation technique of food that inactivates harmful pathogens and vegetative spoilage microorganisms. HPP is an alternate, non-thermal food processing method, wherein the food is subjected to a very high pressure range from 100 -800 MPa (1MPa = 145.03 Psi or 10 Bar). HPP is also referred as High hydrostatic pressure processing, Ultra high-pressure processing, Hyperbaric processing and Pascalization (Rao et al., 2014).

Milk is considered to be a perfect food which contains almost all the macroand micronutrients required for normal body growth. However, it is highly perishable in nature which poses problem during its storage. It gets spoiled within few hours after milking if not processed or cooled immediately. It can also be a source of several disease causing microbes as it is conducive for the growth of microorganisms.

The Indian economy, which meets its own dairy demands, has reached a turning point with its bovine breeding initiatives. The viability of this enterprise, however, depends largely on completing as many successful artificial insemination cycles at a cattle farm. Estrus behaviour observation is used to determine when artificial insemination should take place. But since not all cattle exhibit substantial estrus symptoms, insemination results are not always reliable. An estrus detecting instrument known as an Electronic Nose (eNose) is created to address these issues.

The world’s greatest source of cow and buffalo diversity is India, and countries like Australia, South Africa, Latin America, and the USA employ Indian cattle breeds to grow their livestock industries. The number of cattle in the nation represents 17% of all cattle in the world. 22-25% of the Indian cow population is made up of the best native milk, draught, and dual-purpose animal genetics. The majority of local cattle breeds are excellent drafters. Native livestock breeds are clearly superior in their use of subpar feed, have better heat tolerance, and greater resistance to tropical diseases.

In India, the Ministry of Health and Family Welfare (MOH&FW), dealing with the subject of food quality and safety at the national level, has been designated the nodal ministry for maintaining food standards. The instrument for ensuring food quality and safety at the national level is through a legislation titled Prevention of Food Adulteration Act, 1954 (PFA) and Rules made there under as amended from time-to-time.

All across the world, the dairy industry is facing numerous problems and changes. Dairy development has received a considerable boost from factors including access to technical advancements, improvements in infrastructural facilities, and the development of modern marketing insights. It is interesting to see how the dairy sector is changing globally.

India has an enormous wealth of cattle, which is crucial to the development of the socioeconomic status of millions of rural households as well as the country’s economy. In addition to giving millions of people access to lowcost nutritious food, this industry plays a significant role in boosting family incomes and creating gainful employment in rural areas, notably among landless laborers, small and marginal farmers, and women.

The world’s largest population of milch cattle lives in India. In 2020, there were almost 906 million tones of milk produced worldwide, with India being the top producer with a 22 percent share. Annual Growth Rate of milk production in 2019-20 in India was 5.69 per cent. Highest growth rate in milk production was found to be 14.31 per cent in Karnataka. In 2021, the Indian dairy market had a value of INR 13,174 billion.

The cradle-to-grave dairy supply chain includes a number of operations that result in greenhouse gas (GHG) emissions related to milk production. These activities broadly include the use of agricultural inputs like fertilizers, fossil fuel for the production of animal feed and fodder, processing concentrates in cattle feed plants, enteric fermentation, manure management, and post farm gate activities like processing milk at dairy cooperative societies, milk unions, its conversion into different milk products, and their distribution through retail supply chains.

Operation Flood and dairy cooperatives became India’s greatest rural employment programme, enabling the modernization of the dairy industry to a point from where it may take off to fulfill the need for milk and milk products within the nation as well as chances on the international market. An unorganized sector handles 80% of the milk a rural producer produces, and an organized sector handles the remaining 20%. It has been discovered that dairy cooperatives significantly contribute to reducing rural poverty by increasing rural milk production and marketing.

Letting the cow to release its milk so the dairy farmer can extract it for personal use or to sell is the procedure of milking. Consequently, it is not totally a natural process. The natural process must be manipulated by the dairy farmer to his advantage. Therefore, in order to control the natural process, one must first understand it. The process of producing milk from blood-derived basic materials by alveolar cells and storing it in the alveolar cavity is known as milk secretion. Only when the alveolar cavity is full and the pressure on the alveolar cells prevents further secretion does this continuous process come to an end.

Animals need to function efficiently during reproduction in order for a dairy production system to be sustainable. This is necessary to produce milk, the primary product of interest, as well as to supply replacement animals. From the perspective of reproduction, delayed puberty, lengthy calving intervals, limited productive lives, and high calf mortality are the key variables that cause economic losses. Reproductive management’s proportionate contribution to the effectiveness of dairy production systems varies greatly.

Dairy farmers hope to have better milking cows than those in their current herds. A solid foundation is necessary for every farming operation. Undoubtedly, the dairy farm documentation system is among the most important facets of farm management. Tracking each cow’s productivity within a herd and assisting breed development generally are the major objectives of a dairy farm tracking system. This information could be referred to as production data.

By minimizing the harsh climate effects like heat, cold and rain, it is crucial to offer the proper circumstances for raising dairy cows in the tropics. Stress can be reduced through the design of the farm and the dwelling. Controlling the environment lowers the risk of stress and disease, improves management, and increases milk production.

Dietary fat, which is not degraded in upper part of digestive tract (rumen) of animal, but gets digested in lower alimentary tract is known as bypass fat. Rumen protected fat is another name for this bypass fat. Immediately following calving, dairy animals lose a significant amount of energy through milk, although their feed intake is decreased. Animals continue to have negative energy balances as a result.

For the purpose of passing into the lower gastrointestinal tract and supplying the cow with vital amino acids, dairy bypass protein (UDP – undegraded protein) is a type of animal or plant-based protein that resists breakdown in the rumen. More metabolizable protein is made available by avoiding the rumen, assisting the cow in meeting her needs for amino acids for milk production and its constituent parts.

In terms of their contribution to human nutrition, the dairy industry is very important. Since dairy farming has the ability to offer meaningful selfemployment and stable income, it is a dependable source of subsistence for the rural poor. Additionally, it requires less labour and cash to maintain. Dairy cattle feeding directly affect the animals’ ability to reproduce and produce. Therefore, the addition of a mineral mixture is necessary to maximize the animal’s productivity and milk output.

The proper supply of feed and fodder is a requirement for the growth and development of the animals. Over the past few decades, there has been a considerable growth in milk production, with India now producing the most milk in the world (187.7 million tones in 2018–19). The main causes of India’s low milk yield are extrinsic (poor nutrition/feed management) as well as intrinsic (low genetic potential).

Herd management practices in the handling, nutrition, milking techniques, sanitation, and housing of dairy animals play key roles in predisposing the individual animal as well as herds to diseases. The key factor influencing the reduction of health issues are the decisions made by herd managers or animal owners.In order to maximize output and profitability through disease control, herd management approaches in conjunction with a veterinarian programme can be most beneficial.