eChapters

Introduction

Small and medium-sized industrial units, or small scale industrial (SSI) units, are generally referred to as SMEs, according to the Ministry of Micro, Small and Medium Enterprises. The MSMED Act of 2006, which was updated in 2020, serves as the foundation for the definition of SMEs in India.

• Micro manufacturing and services units: Up to Rs. 1 crore in investments and up to Rs. 5 crore in revenue.

• Small manufacturing and services units: Up to Rs. 10 crore in investments and up to Rs. 50 crore in revenue.

• Medium manufacturing and services units: Up to Rs. 50 crore in investments and up to Rs. 250 crore in revenue.

Small and Medium-sized enterprises (SMEs) constitute the backbone of the global economy. SMEs not only provide major contributions to national GDPs, but they also play an important role in job creation, innovation, and regional development. In India, SMEs account for about 30% of the country’s GDP and employ over 110 million people, making them an important part of the economic structure.(FICCI,2023).

Introduction

Entrepreneurship is crucial for economic development, especially during crises when job losses push people to find alternative livelihoods, often turning to entrepreneurship as a means of survival. Its impact is highly beneficial, promoting growth, wealth creation, and better living standards. In developing countries like India, entrepreneurship offers a path to holistic development, particularly for disadvantaged groups traditionally dependent on agriculture. Uplaonkar and Biradar (2015) noted that entrepreneurship development effectively generates employment, increases income, reduces poverty, improves health, and ensures food and nutritional security. Recognizing the importance of entrepreneurship, stakeholders in the agricultural sector now see its positive impact on business sustainability. Verhees et al. (2011) argued that promoting entrepreneurship in agriculture can significantly reduce poverty and boost rural prosperity, directly addressing socio-economic challenges in rural areas (Bhooshan and Ranjith, 2022). Therefore, supporting the entrepreneurial ecosystem in rural sectors through programs and schemes is essential.

Introduction

Entrepreneurship, known for its ability to drive innovation, economic development, and societal change, is a dynamic force that plays a crucial role in global advancement. However, the path of an entrepreneur is not a straightforward journey from concept to achievement. Instead, it is a complex process influenced by numerous factors, which together form the entrepreneurship development cycle. Gaining a deep understanding of this cycle, along with the entrepreneurial environment and the challenges entrepreneurs face, is essential for cultivating a thriving entrepreneurial ecosystem.

The entrepreneurship development cycle refers to the various stages that entrepreneurs go through as they turn ideas into successful ventures. The cycle starts with ideation, where entrepreneurs identify opportunities and develop innovative solutions to address unmet market needs or problems. This is followed by a feasibility assessment, during which entrepreneurs assess the potential of their ideas, considering market demand, competition, and the resources needed. After confirming the viability of their concepts, entrepreneurs move on to resource acquisition, gathering the capital, talent, and infrastructure necessary to bring their ideas to life. The subsequent phases include launching the venture, establishing a market presence, and pursuing growth through strategic decision-making and adaptability to shifting market conditions.

Introduction

Meaning of Entrepreneur

The term “entrepreneur” is derived from the French verb entreprendre, which means “to undertake.” Historically, its meaning has evolved. In the early 16th century, it referred to individuals involved in engineering tasks like construction. By the 18th century, the term came to describe those engaged in economic activities. Over time, the concept of an entrepreneur shifted from someone participating in military ventures to someone actively involved in business and economic pursuits. Today, an entrepreneur is defined as an individual who innovates, organizes resources, and takes on risks to create new ventures or products with the goal of generating profits.

Richard Cantillon was the first who proposed the term “entrepreneur is an agent who buys factors of production at certain prices in order to combine them into a product with a view to selling it at uncertain prices in future.”

Introduction

Entrepreneurship has become a major driver of economic development in recent years. It involves starting new businesses or revitalizing existing ones, often through recognizing and seizing new opportunities. Entrepreneurs are individuals who organize, manage, and take on the risks of a business venture with the aim of making a profit. They identify market opportunities and introduce new technologies and organizational improvements. Entrepreneurs constantly innovate and create value from these opportunities. According to Hisrich (1990), entrepreneurs are proactive and creative individuals who can organize resources and manage risks and failures to succeed.

To grow as an entrepreneur, it is essential to develop and refine key entrepreneurial traits. These traits shape an entrepreneur’s skills and abilities, aligning with their motivations and guiding them to drive their business forward. Entrepreneurial characteristics are a mix of personal, social, and environmental factors that influence a business ecosystem. These traits play a key role in shaping a company’s goals and strategies, ultimately affecting its performance.

Concept of Entrepreneurship

Entrepreneurship can be defined as the propensity of mind to take calculated risks with confidence to achieve a pre-determined business or industrial objectives. That points out the risk-taking ability coupled with decision making. The word ‘entrepreneurship’ typically means to undertake. It owes its origin to the western societies. But even in the west, it has undergone changes from time to time. In the early 16th century, the term was used to denote army leaders. In the 18th century, it was used to denote a dealer who buys and sells goods at uncertain prices. Towards 1961, Schumpeter, used the term innovator, for an entrepreneur. Two centuries before, the concept of entrepreneurship was shady. It is only in the recent years that entrepreneurship has been recognized widely all over the world like in USA, Germany, Japan and in the developing countries like ours. Gunnar Myrdal rightly pointed out that Asian societies lack entrepreneurship not because they lack money or raw materials but because of their attitudes. Till recently, in the west, the entrepreneurship is mainly an attribute of an efficient manager. But the success achieved by entrepreneurs in developing countries demolishes the contention that entrepreneur is a rare animal and an elusive character. In India the definition of an entrepreneur being the one who undertakes to organize, own and run a business has been accepted in a National Seminar on entrepreneurship organized in Delhi in 1975. Still there has been no consensus on the definition of entrepreneurship and qualities of entrepreneurship.

Introduction

Globalization, as defined by the International Monetary Fund (IMF), refers to the increasing economic interdependence among nations, characterized by rising volumes and diversity of cross-border transactions in goods and services, international capital flows, and the more rapid dissemination of technology worldwide.

Key Characteristics of Globalization

1. Expansion opportunities for businesses globally.

2. Elimination of distinctions between domestic and international markets.

3. Freedom to buy and sell products and services across any country.

4. Production and distribution facilities are established based on economic feasibility at optimal locations worldwide, rather than national priorities.

5. Product planning and development are influenced by global market conditions.

6. Sourcing raw materials from across the globe to maximize efficiency.

7. Global strategies, organizational structures, cultures, and management practices.

8. Viewing the world as a unified market.

Introduction

In simple terms, business refers to buying and selling of products. Business would define itself as an organized effort through companies aimed at supplying consumers with products and services in a return for monetary compensation. However, this simplistic definition is insufficient to assess the impact of the environment on modern international business operations. For easy understanding, modern business can be explained as a complex field of industry and commerce involving activities associated with both production and distribution. These activities cater to the wants and desires of society at one extreme and provide profit for business firms at the other.

Indian leaders, particularly the first Prime Minister Dr. Jawaharlal Nehru, who had conceived the theory of five year plans, had regarded high economic growth and higher income among poor population as the foremost objectives before the newly independent nation. To achieve these objectives, government was made to play a very important role, and since 1951, a series of five year plans have been the driving forces behind the economic prosperity of the country. Despite of experiencing growth in 1950s, it was still lower than that of many Asian countries. Between 1951-79, the Indian economy had grown at an average rate of 3.1 percent a year at constant prices. Growth in the industrial sector had been about 4.5 percent a year, while for the agriculture sector, growth was 3.0 percent over that period. Many of the political leaders associated with the movement were also in support of a socialistic development pattern. They favored state interference in the economy. They encouraged state ownership of core industries.

Introduction

The gut microbiota is the complex and dynamic community of microorganisms that includes various bacteria, fungi and other microorganisms, proliferate and lead a symbiotic/mutual relationship with the host system. These microorganisms protect the host against pathogens by unique mechanisms, helps to maintain shape and strengthen the intestinal epithelium and boost immune response. Diverse bacterial metabolites can immigrate the host bloodstream and other parts of the body via various channels in humans and animals, such as gut-microbiota-brain, gut microbiota-skin, gut-vagina, gut liver, gut-bones etc. However, many aspects of modern lifestyle would alter the gut microbiota and lead to numerous diseases including inflammatory bowel disease, diabetes mellitus, obesity, and metabolic syndrome. Probiotics and other beneficial microbes exert many beneficial effects on their hosts by modulating the gut microbiota. While the term probiotic is reserved only for well-characterised strains with clinically proven health benefits, other beneficial microorganisms include, organisms responsible for the fermentation of foods viz. yogurt, kefir, kombucha, kimchi and many others that have been shown to have distinct health benefits. Interestingly, major group of these beneficial organisms belong to lactic acid bacteria.

Introduction

Lactic acid bacteria (LAB) are characterized as Gram-positive cocci or rods, non-aerobic but aerotolerant, able to ferment carbohydrates for production of energy and lactic acid. Lactic acid bacteria includes different major genera: Lactococcus, Enterococcus, Lactosphaera, euconostoc, Melissococcus, Oenococcus, Pediococcus, Streptococcus, Tetragenococcus, Vagococcus and Weissella. Other genera are Aerococcus, Microbacterium, Propionibacterium and Bifidobacterium. Through carbohydrate fermentation along with lactic acid, various organic acids are produced due to which there is a fall in pH level and creates unfavorable environment or inhibits growth of some undesired microorganisms. The low pH makes organic acids lipo -soluble, allowing them to invade into cytoplasm of pathogens by rupturing cell membrane. LAB are generally regarded as safe (GRAS), and have a significant role in the preservation of foods and fermented products. Under controlled conditions they can be used as natural competitive microbiota or as specific starter cultures. Antagonistic substance like bacteriocins are produced from lactic acid bacteria that inhibits the activity and growth of other bacteria and these bacteriocins are active against various pathogens in minimum concentration. These are antimicrobial peptides modified by post -translational mechanisms and exported to the extracellular medium. Till date about 239 bacteriocin producing LAB genome and 789 putative bacteriocin producing genes cluster are stored in the public databases. Many gram positive and gram negative bacteria in the environment produce bacteriocins.

Introduction

Let food be thy medicine and medicine be thy food- the age-old quote by Hippocrates, is certainly the tenet of today. Microbial cultures have been used for millenniums in food and alcoholic fermentations and in the past century have undergone scientific scrutiny for their ability to prevent and cure a variety of diseases. Later it became popular in improving human and farm animal nutritive values and recently in aquaculture also. The growing interest in the understanding the role of food in human health has moved from its primary role as a source of energy to the subtle action of biologically active food components on human health. Henceforth functional foods or nutraceuticals are of great demand in present time. The growing scientific evidence suggests that the food supplements containing beneficial bacteria can provide an array of benefits to the host. Now a day’s probiotic fermented foods are in great demand due to the awareness of beneficial aspects of probiotics among people. Several industries focused on the improvement of conventional fermented foods scientifically in order to fetch good market for their products. Particularly, animal feed companies and researchers have been looking for alternative products and strategies that can help to maintain animal gut health in order to prevent or reduce the prevalence of pathogens in the food chain. After understanding the mechanism and beneficial aspects of probiotics in detail at molecular level in earlier chapters, now it is relevant to discuss the applied part of probiotics. In this chapter, commercial importance of probiotics and probiotics derived products at industrial level are emphasized.

Diversity of gut microbial community

The human microbiota (the collection of microbes that live on and inside us) consists of about 100 trillion microbial cells that outnumber our ‘‘human’’ cells 10 to 1, and that provide a wide range of metabolic functions that we lack. It has been estimated that they encode 100-fold more unique genes than our own genome. Furthermore, the gut microbes contribute to energy harvest from food, and changes of gut microbiome may be associated with bowel diseases or obesity. To understand and exploit the impact of the gut microbes on human health and well-being it is necessary to decipher the content, diversity and functioning of the microbial gut community. If we consider ourselves as superior organisms encompassing these microbial symbionts, by far the majority of genes in the system are microbial. In this sense, completing the human genome also requires the characterization of the microbiome (the collection of genes in the microbiota).

Methods of counting viable and non-viable bacteria in gut

Before the advent of genome sequencing and genome-wide analysis method, our knowledge of molecular characteristics of diverse gut microbial population was extremely limited. To obtain an estimate of cultivability, microbial ecologists generally compare microscopic counts with total viable counts. The total viable count made on a nonselective agar based medium estimates the number of colony forming units (cfu) per gram of sample. The finding that total viable counts are typically lower than total microscopic counts was thought to be due to the number of dead cells. Indeed, dead bacteria in feces may constitute up to one third of the total bacterial community. However, recent nucleic acid based studies indicated that a majority of bacteria in a variety of ecosystems are different from those described in culture.

Introduction

It became clear from the discussions of previous chapters that probiotics perform metabolic functions such as fermenting indigestible dietary residues and endogenous mucus, production of vitamins, and absorption of minerals. They also play key role in intestinal epithelial cell proliferation and differentiation, and the homeostasis of the immune system. Further, probiotics are also known to involve in modifying gut pH, antagonizing pathogens through the production of antimicrobial compounds, competing with pathogens for recept or sites as well as for available nutrients and growth factors, stimulating immunomodulatory cells of the host, and produce a wide range of enzymes such as lactase, amylase, lipase, protease, phytase and so on. The molecular mechanisms of probiotics are largely unknown and opens avenue for research in probiotics. Moreover, current research in probiotics and health consciousness in the public lead to an increased interest towards probiotics from the public, researchers, governmental organizations (such as the WHO/FAO) and medicine and food companies. In this chapter, application of probiotics in treating various diseases and disorders are discussed.

Introduction

Cell surface components such as adhesins, polysaccharides, and proteins play major roles in the adherence of probiotic bacteria to the intestinal epithelium, interactions that might lead to pathogen exclusion and immunomodulation of host cells. Probiotic bacteria have the ability to produce exocellular polymers called Exopolysachcharides (EPSs). These exocellular polymers present in the surface of many lactic acid bacteria, including Lactobacilli.The adhesive properties of lactobacilli are directly linked to their surface properties which are influenced by the structure and composition of their cell wall. The cell envelope of lactobacilli, bacteria, is composed of the bilipidic plasma membrane with embedded proteins encompassed by the cell wall. The bacterial cell wall consists of a thick multilayered sacculus made of peptidoglycan (PG) with teichoic acids (wall teichoic acids (WTA) and/or lipoteichoic acids (LTA)), exopolysaccharides (EPS), proteinaceous filaments called pili, and proteins that are anchored to the cell wall through different mechanisms (Figure 1).

Some species of lactobacilli possess an additional paracrystalline layer of proteins surrounding the PG layer, referred to as the S layer. These macromolecules together may play crucial roles in determining species and strain-specific characteristics of lactobacilli by influencing host-microbe interactions and microbial adaptations to the changing host environment. Several studies implicate that cell surface components, either individually or collectively, involve in host-microbe interactions. Lactobacilli beneficially affect the host in many ways by producing abundant variety of EPSs through the activity of extracellular glycosyl transferases. Approximately 30 species of lactobacilli are described as EPS producers. Among them, the best known are L. casei, L. acidophilus, L.brevis, L. curvatus, L. delbrueckii bulgaricus, L. helveticus, L. rhamnosus, L. plantarum, L. johnsonii, etc.

Introduction

Preceding chapters discuss the essentials of probiotics, their role in maintaining gut health in combination with prebiotics and significance of systematic classification of probiotic isolates. Further, to draw an idea how probiotics function in the gut, it is essential to know the interaction of probiotics with other microorganisms in the gastrointestinal milieu - the site of their probiotic action. Knowledge in the ecology of probiotic microorganisms in host environment such as gut gives an ideal picture of their functioning manner and the factors (both host and microbial) responsible for it.

Study of microbial ecology in the gastrointestinal tract is very difficult since gut harbors highly diversified microflora that varies not only at species level but also at individual level. The diversity of gut microflora in two healthy individuals of same species may exhibit variations in their composition and it may be influenced by geodemographic factors also. In this regard, the present chapter deals with the study of ecology of Lactobacillus- most commonly used probiotic bacteria in the gastrointestinal tract which in turn helps in framing selection criteria for screening of potential probiotics.

Recent studies on interaction between human and gut microflora discussed that co-evolution between microorganisms and their host have contributed for the development of a very complex microbial diversity in the gut. Highly diversified microflora in the human gut is a consequence of natural selection at two levels, top-down and bottom-up selection pressure. Hierarchy theory expounds that higher levels compel lower levels for possible organizational solutions i. e. organisms at the higher levels govern the structure or population dynamics of an ecosystem while on contrary lower levels control the structure of ecology when bottom-up level is the selection pressure. In case of interaction between human and gut microflora, being host human is at higher level in the hierarchy for selection and as mentioned above, top down selection pressure is driven by higher levels; host selects the beneficial microorganisms for functional redundancy leading to the development of a microbial community with divergent microbial divisions. Chances for lateral gene transfer also thought to influence the selection process and there by structure of the microbial community. On the other hand, competition among the members of host selected microbiota exert bottom up selection pressure resulting in the development of highly specialized genomes with functionally prominent set of genes. Natural selection at host level obviously favors functionally constant microbial communities which benefits the host with functional redundancy encoded in their genomes. Mean while bottom-up cell-level selection pressure aspire to favor niche specialization to avoid competition between the members of microbial community. In order to be a part of specialized niche, to become more potent microorganisms adapt a particular set of abiotic and biotic factors within the habitat by acquiring new sets of genes essential for their survival and thus escape the competition from decrepit microbial species.

Introduction

From earlier chapters, it is evident that Lactobacilli and Bifidobacteria are the two important genera that have potent applications in food microbiology and human nutrition due to their role in food and feed production, preservation and also due to probiotic potentialities. Most of the bacterial species are still unknown indicating that the identification and classification of bacteria is a hard task since the beginning of microbiology and consequently our knowledge about bacterial ecology is very low. Mean while the expectations about novel bacterial products are high. Therefore, bacterial identification has become a growing field of interest in microbiology. Since, Lactobacilli and Bifidobacteria genera include a large number of species and strains exhibiting important properties in an applied context, chiefly in the area of probiotics as food and therapeutics, an updated list of species belonging to these two genera, their phylogenetic relationships and other relevant taxonomic information are of great significance. The classification and identification of a probiotic isolate at strain level point out its typical habitat and origin. Moreover, the species, or genus name, may also indicates the strain’s safety and technical applicability for its use in probiotic products. Taxonomy helps to place a newly identified organism in a relevant group and enables the scientific communication. Further it facilitates the construction of databases that can be used for rapid identification of organisms at laboratory level easily. Thus, taxonomy of Lactobacilli and Bifidobacteria helps to study these organisms scientifically and allows characterizing and arranging them in an orderly manner which promote the scientific community to explore the benefits from those organisms for the betterment of mankind.

Introduction

Probiotics, prebiotics and synbiotics are interrelated terms. Probiotic is discussed in detail in previous chapter. In this chapter, prebiotics and its effect on gut along with probiotics are emphasized. Prebiotics can be defined as non digestible food ingredients that stimulate the growth and/or activity of bacteria in the digestive system in ways claimed to be beneficial to health. They were f irst identified and named by Marcel Roberfroid in 1995. He defined prebiotic as a selectively fermented ingredient that allows specific changes, both in the composition and/or activity in the gastrointestinal microflora that confers benefits upon host well-being and health. Further, FAO defined prebiotic as “a non-viable food component that confers a health benefit on the host associated with modulation of the microbiota.”Whereas the term synbiotics refers to a product that contains both probiotics and prebiotics in the form of synergism. A synbiotic has been defined as “a mixture of prebiotics and probiotics that beneficially affect the host by improving the survival and implantation of live microbial dietary supplements in the gastrointestinal tract, by selectively stimulating the growth and activating the metabolism of one or a limited number of health promoting bacteria, and thus improving host welfare”. As described above, the combined use of prebiotics and probiotics is often described as synbiotic, but the United Nations Food & Agriculture Organization (FAO) recommends that the term “synbiotic” be used only if the net health benefit is synergistic.

The term probiotic means “for life” and it is currently used to refer bacteria that are associated with beneficial effects for humans and animals. The credit for first observation made on the positive role of some selected bacteria was attributed to Eli Metchnikoff, the Russian born Nobel Prize recipient, worked at the Pasteur Institute in the beginning of the last century. Metchnikoff suggested that - “the dependence of the intestinal microbes on the food makes it possible to adopt measures to modify the microflora in our bodies and to replace the harmful microbes by useful microbes”. Interim Henry Tissier, a French paediatrician, observed that children with diarrhea in their stools had a low number of bacteria which are characterized by a peculiar Y shaped morphology. On the contrary, these “bifid” bacteria were abundant in healthy children. Based on these observations, Tissier suggested that “bifid” bacteria could be administered to patients with diarrhoea to help restore a healthy gut microflora. These two scientists were the first to give scientific intimation for the use of probiotics when even the term probiotics was not coined. Later in 1965, the term probiotics is coined by Lilly and Stillwell to describe substances produced by one microorganism, that stimulate the growth of another. Subsequently, Parker defined probiotics as organisms and substances that contribute to intestinal balance. Further, Fuller (1989) improved the Parker’s definition of probiotics as “live microbial feed supplements which beneficially affect the host animal by improving its intestinal microbial balance”. Havenaar and Huis in’t Veld (1992) extended the probiotic definition with the description: “A viable mono or mixed culture of microorganisms which, applied to animal or man, beneficially affects the host by improving the properties of the indigenous microflora”. At the end of the millennium, Salminen and his coworkers (1999) proposed the following definition: “probiotics are microbial cell preparations or components of microbial cells that have a beneficial effect on the health & well- being of host”. Whereas FAO/WHO (2001) defined probiotics as “living bacteria that, when administered in adequate amounts, confer a health benefit on the host”. This definition for probiotics is widely accepted.

The contribution of dairy sector in the Indian economy has tremendous impact on the country’s GDP. India being the world’s highest milk producing country, dairy industry has a major role in changing the twin problem of the nation that is poverty and unemployment. Dairy sector also involves the under privileged group, especially the dairy farm women. There are many success stories regarding the life changing experiences gained by the farm women, who had taken dairy farming as a means to sustain their livelihood. In the area of gender equality, dairy sector has proved to be a women centric enterprise. Most of the activities pertaining to dairy farming are mostly carried out by the farm women. The dairy farming activities carried out by the farm women are tedious and in major cases their contribution remains unnoticed. They are usually carrying out activities, where most of their energy is drained, resulting in drudgery. Matters related to decision making, their opinion about adopting new technologies, starting an enterprise are not given importance by their family members. This leads to a greater setback in the emergence of success of dairy sector. Therefore, empowerment of farm women in dairy sector is one of the important criteria for the dairy industry to reach a greater height and for overall development of nation.

This chapter presents the analysis, interpretation and discussion of the collected data from the study area in an effort to attain the objective of the study. The data collected were analyzed with appropriate statistical test and presented in a systematic format.

The results of the study are presented under the following subheadings:

4.1 Distribution of respondents according to different socio-economic variables

Basic information of the respondents under the study has been given according to the socio-economic characteristic of the respondents. Variables like age, education, marital status, family size, family type, milk production per day, social status, farming experience, herd size, land holding and annual income are presented in the following table 4.1

Research methodology gives detailed justification about the research methods and procedures carried out in the present study. The present study focuses on the women’s involvement in the dairy farming and their empowerment. Keeping in view the specific objectives of the study and various aspects relating to these objectives the methodological framework has been formulated. The sub topic under the research methodology is as follow:

3.1 Locale of the study

The present study entitled “Retrospective Multidimensional Study on Magnitude of Participation of Women in Dairy Farming in East district of Sikkim” was carried out in the villages of East district of Sikkim. Sikkim is the second smallest states in India, with an area of 7,096 sq. km in size. The meaning of Sikkim is “The valley of rice”. It lies between 27 º33' 00" N and 88 º30' 00" E. The average annual temperature of the state is 18 ºC (64 ºF) and annual rainfall is 2739 mm. The state is situated in the Eastern Himalayas and it shares its border with countries like Tibet, Nepal and Bhutan. Mt Kanchendzonga (8585m) which is the third highest peak in the world is situated at Sikkim. According to 2011 census, population of state was 610,577 with male population of 323,070 and female population of 287,507. The literacy rate in the state is 81.40 percent. The density of the state is 86 people per km2 (Census,2011). The cattle population in the state was 264,860 in number (BAHS,2019). The state has four district- East, West, North, and South (Registrar General of India, 1989).

Research being a continuous process for a scientific study, previous study provides basis for the research. The review of literature is necessary for developing conceptual frame work and suitable design for the study. It assists the researcher to keep his work going in right direction. The available literature related to the present study has been reviewed under the following heading:

2.1 Differential role accomplished by women dairy farmers from different social strata

Toppo et al. (2004) examined the participation of women farmers in dairy occupation in Anand district of Gujarat and their result revealed that 18.34 percent of the farm women were mostly engaged in activities related to artificial insemination, majority of the respondent (68.34%) participated in cleaning of the shed, while 25.00 percent of the farm women practised manure preparation and 95.00 percent of the respondent were not involved in preparation of gobar gas mixture.

Badole (2006) reported that majority of the responded had low socio-economic status.

India is one of the most populous country in the world with a population of 1.26 billion (Census,2011). Of which women constitute about 49 percent of the population. Women act as a significant factor in development of the nation. Women from all sphere of life are contributing meaningly towards the prosperity of the nation. Especially, the rural farm women who constitute nearly half of the total rural population. The rural women are the mainstay of Indian agriculture. Their contribution towards agriculture development is worthy of praises. They are capable of accomplishing sustainable development in all the various activities which they have carried out and shown interest. Women are the essential part of society. For a society to develop, women should be given equal importance as the male counterpart. Nurturing of society is possible only if women are nurtured with equivalent opportunities and progress. Indian society is still male dominant, which accounts for low reputation of women. Women are treated as less competent human being. Through passing of time there have been acknowledgement by the Constitution of India on the part of importance of women as a productive humane resource for development of nation. Many measures have been adopted for the gender equality. This paved the way for advancement of empowerment of women.

Vitamins are naturally occurring, vital nutrients that are needed in small amounts. They are important for bone and tissue health, wound healing, growth and development, immune system function, and other biological processes. These vital organic substances serve a variety of biochemical purposes. Vitamins, like minerals, are not biosynthesized by the body. Consequently, in order to maintain our health, we must obtain them from the food we eat or, in rare circumstances, supplements.

Vitamins in Milk

Together with water- soluble vitamins like vitamin C and B complex and provitamins, milk also contains a variety of fat- soluble vitamins like A, D, E, and K. Some of these vitamins do, however, have very low concentrations.

Vitamin A (Retinol)

Plant foods contain carotenoids, which are the precursors of vitamin A, whereas animal foods provide preformed vitamin A in the form of retinyl esters.

The primary constituents of milk are water, fat, protein, sugar (lactose), minerals, vitamins, and enzymes. Milk is a thin emulsion made up of two phases: A continuous aqueous colloidal phase and a dispersed phase of oil or fat. It is an opaque white liquid containing fat in an emulsion, protein, and a few minerals in a colloidal suspension, and lactose along with soluble proteins and a few minerals in a true solution. The physical characteristics of milk are comparable to those of water, but they are altered by the degree of dispersion of the colloidal and emulsified components as well as the presence of different solutes, such as salts, lactose, and protein, in the continuous phase.

Physical Appearance, Color and Optical Properties

Fat globules and casein micelles scatter light, giving the appearance of turbid and opaque milk. The way that molecules scatter light has an impact on optical characteristics. When the wavelength of light and the particle’s magnitude coincide, light scattering happens. Consequently, light with shorter wavelengths is scattered by smaller particles and vice versa. Because casein micelles scatter blue light more than red light at shorter wavelengths, skim milk appears slightly blue. Cow’s milk has a creamy color because of beta- carotene, a carotenoid precursor to vitamin A. The greenish tinge in whey is due to the presence of riboflavin. Turbid and opaque due to scattering of light mainly by protein micelles and fat globules.

Electrophoresis

The mobility of ions in an electric field is the basis for the separation method known as electrophoresis. Ions that are negatively charged move toward a positive electrode, while those that are positively charged move toward a negative electrode. Usually, one electrode is biased either positively or negatively, and the other is at the ground for safety reasons. Ions can be distinguished from one another based on their size, shape, and overall charge, which determine their migration rates.

Poly Acrylamide Gel Electrophoresis (PAGE)

Introduction

Electrophoresis is an electrochemical process in which substances with a net electric charge migrate under the influence of an electric current. This migration occurs in an agar gel or a liquid- filled buffered matrix such as cellulose acetate. Positively charged substances travel toward the cathode (negative electrode), while negatively charged substances go toward the anode (positive electrode). Different substances move at different rates depending on their charge. This movement is called electrophoretic mobility.

Proteins are charged molecules. They have a positive as well as a negative charge. Among proteins, particularly caseins, they have different contents. The charge density also varies depending on the molecular weight. Proteins have a net negative charge at a pH above 4.6, and hence they move towards the anode (positive electrode) when subjected to an electric field under alkaline conditions. Due to differences in the charge of individual protein fractions, their migration rates also differ and are hence separated into distinct bands. These bands are stained with the dye amido- black, destained with 7% acetic acid, and observed for the electrophoretic pattern.

Food adulteration is a global issue, and because there are insufficient regulations and oversight, developing nations are more vulnerable to it. But in many nations, food adulteration has gone unnoticed. Contrary to popular assumption, milk adulterants can, regrettably, present major health risks that result in fatal diseases. Modern milk adulteration involves increasingly complex techniques, necessitating state- of- the- art research to identify the adulterants. Because milk contains so many nutrients that are needed by both adults and infants, it is referred to as the ideal food. In terms of protein, fat, carbohydrates, vitamins, and minerals, it is among the best sources. Regretfully, milk adulteration is a very common problem worldwide. Potential causes could be the gap between supply and demand, the perishable nature of milk, the low purchasing power of the consumer, and the absence of appropriate detection tests. Because there is insufficient oversight and inadequate law enforcement, the situation is substantially worse in developing and underdeveloped nations. While quantitative detections of adulterants in milk are varied and complex, qualitative detections can be carried out simply using chemical reactions. The type of quantitative detection methods used depends on the milk’s adulterants.

Typical Adulterants in Milk

The addition of vegetable protein, milk from different species, whey addition, and watering are the main adulterants in milk that are known to be economically motivated. There is no serious health risk associated with these adulterations. Some adulterants, though, are too dangerous to ignore. Urea, formalin, detergents, ammonium sulfate, boric acid, caustic soda, benzoic acid, salicylic acid, hydrogen peroxide, sugars, and melamine are a few of the main adulterants in milk that have been shown to have serious negative health effects.

The terms “cultured dairy foods,” “cultured dairy products,” and “cultured milk products” are other terms for fermented dairy products. Lactobacillus, Lactococcus, Leuconostoc, and other lactic acid bacteria, along with yeasts in certain products, are used to ferment milk to make these dairy products. The product’s shelf life is extended through fermentation, which also improves the product’s taste and milk’s digestibility. There is a wide variety of fermented milk products available, each with unique flavors and attributes. The following categories apply to fermented milk products, based on the type of fermentation.

Changes in Nutrients Resulting from Fermentation

Fermentation lowers the lactose content in fermented milk products; however, if the starting milk is concentrated or fortified, the finished product’s lactose content may be higher than that of liquid milk.

The composition and content of protein and total amino acids are comparable to those of raw milk. Casein degradation is small compared to cheese, accounting for not more than 1% of the total protein. While some products of casein degradation are used by the starter microorganisms, others build up as free amino acids in the fermented kinds of milk. The amounts of free amino acids that build up differ depending on the species. Generally, proline is increased in large quantities, followed by serine, alanine, valine, leucine, and histidine. As a result of continuous starter culture activity, the level of free amino acids increases during storage. A limited degree of lipolysis occurs during the manufacture of fermented milk. The amount varies depending on the type of species, but the pattern and concentration of free fatty acids are both impacted. During fermentation and the breakdown of amino acids, trace amounts of volatile fatty acids are also produced.

Being an essential component of the rich Indian past, traditional dairy products and a sophisticated classification of confections have played a significant role in the Indian economy. Depending on how it is processed, it has significant social, cultural, and economic significance. These products have been developed over a long period using a combination of experience, traditional wisdom, and culinary skills. India is emerging as the highest milk- producing country in the world with an annual production of 221.1 million tons in the year 2021–22 (Source: Basic Animal Husbandry Statistics, MoFAHD, DAHD, GoI). An estimated 50% of the milk produced in India is used to make a range of traditional dairy products, including paneer, dahi, ghee, khoa, and shrikhand. The product is not only well- established in the Indian market, but it also has significant export potential due to the large Indian diaspora living all over the world.

All native milk products that have developed over time using locally accessible technologies are referred to as traditional Indian dairy products or Indian indigenous milk products. The Indian masses’ economic, social, religious, and nutritional well- being are greatly influenced by traditional dairy products.

Classification of Traditional Indian Dairy Products

Traditional dairy products in India are typically made, packaged, and/or sold using time- honored methods without the use of machinery or meticulous documentation of the procedure. These products do, however, have several technological advantages, including their widespread appeal, reduced cost of production, straightforward manufacturing process, adoption at the rural level or collection center, large profit margin, and abundance of employment opportunities. These products are categorized according to the manufacturing principle.

One of the most consumed and well- known milk products in the Indian subcontinent is ghee. It is a by- product of tradition, prepared both at organized dairy plants and the household level for a well- established market. Ghee is traditionally made from the milk of cows or buffalo. The tetrapyrrole pigments biliverdin and bilirubin are responsible for the distinctive color of buffalo ghee. On the other hand, ghee is golden yellow due to the presence of carotenoids in cow’s milk. Ghee’s composition changes according to the kind and feeding habits of the animal and can differ from place to place and season to season.

In addition to, being a substantial source of fat- soluble vitamins, essential fatty acids, and other growth- promoting components, ghee is a rich source of energy. Its value as food for humans is enormous. Additional attributes related to medicine are present in ghee. Ghee can improve one’s physical and mental attributes and treat eye conditions like ulcers. Ghee is used in religious and ceremonial rites in addition to being consumed.

Composition

Ghee is a complex lipid composed of mixed glyceride which makes up 99–99.5% of its total content. In addition, it has trace amounts of calcium, magnesium, phosphorus, iron, copper, and fat- soluble vitamins, tocopherols, carbonyls, hydrocarbons, carotenoids, free fatty acids, phospholipid, sterols, and sterol esters

The process of making dried milk involves eliminating water from milk, primarily through spray or roller drying. It is regarded as a significant sector of the dairy industry and is made from whole cow, buffalo, or standardized milk. Skim milk powder (SMP) is made from milk after the fat has been removed whereas whole milk powder (WMP) is made from whole cow or buffalo milk

FSSA (2006) Standard Of Milk Powder According to FSSA (2006), Milk powder is the product obtained by partial removal of water from the milk of cows and/or buffalo. The fat and/or protein content of the milk may be adjusted by the addition and/or withdrawal of milk constituents in such a way as not to alter the whey protein to casein ratio of the milk. It shall be of uniform color and shall have pleasant taste and flavor, free from off-flavor and rancidity. It shall also be free from vegetable oil/fat, mineral oil, thickening agents, added flavor, and sweetening agents. It may contain food additives. It shall conform to the microbiological requirements prescribed by FSSA (2006).

Water activity, high storage temperatures, longer than usual storage times, high product moisture content, high processing temperatures, and container headspace are some of the variables that affect the Maillard reaction in dried milk.

Casein and whey proteins are the major proteins present in milk. Unlike casein, whey proteins are heat-labile. Whey protein fractions present in milk are α-lactalbumin, β-lactoglobulin, bovine serum albumin, immunoglobulin, euglobulin, and lactoferrin. β-lactoglobulin and κ-casein interact during heat treatment. When milk is refrigerated for longer than 24 h, it forms γ-casein and protease-peptone fractions.

Commercially, one of the most significant milk products is ice cream. It is a frozen product which is usually made from milk or cream that has been f lavored with fruits like peaches or strawberries, and spices like vanilla or cocoa, or with sugar or a substitute. Stabilizers are sometimes combined with food coloring. Ice cream mix typically contains 10.0% milk fat, 11.0% solids not-fat, 14.0% added sugar, and 0.3% additives like an emulsifier, stabilizer, f lavor, and color substances.

Standard of Ice Cream

According to FSSA (2006), ice cream, kulfi, chocolate ice cream or softy ice cream means the product obtained by freezing pasteurized milk prepared from milk and/or other products derived from milk with the addition of sweetening agents and fruit products, egg products, coffee, cocoa, ginger, and nuts. It may also contain chocolate, and bakery products such as cake or cookies as a separate layer and/or coating. It may be frozen hard or frozen to a soft consistency. It shall be free from artificial sweeteners. It shall have a pleasant taste and smell free from flavor and rancidity. It may contain food additives. It shall conform to the microbiological requirements as given in FSSA (2006).

Concentrated milk products are obtained through partial removal of water which increases the dry matter content of milk. The increase in dry matter content in concentrated milk is proportional to the degree of concentration. The milk concentrate may be prepared from either full-cream whole milk or fat-free skim milk.

Depending on the nature of raw material and other non-milk additives concentrated milks are classified into four broad categories.

1. Sweetened condensed milk

2. Evaporated milk or unsweetened condensed milk

3. Sterile milk concentrate

4. Reconstituted concentrated milk

Sweetened Condensed Milk

One of the earliest dairy products to be produced industrially is sweetened condensed milk. According to FSSA (2006), sweetened condensed milk is the product obtained by partial removal of water from the milk of cow/buffalo with the addition of sugar or a combination of sucrose with other sugars or by any other process that leads to a product of the same composition and characteristics. The fat and/or protein content of the milk may be adjusted by the addition and/or withdrawal of milk constituents in such a way as not to alter the whey protein to casein ratio of the milk being adjusted. It shall have a pleasant taste and flavor and be free from off-flavor and rancidity. It shall be free from any substance foreign to milk. It may contain food additives. It shall conform to the microbiological requirement as laid down by FSSA. The said product shall also conform to the following requirements

Cheese is made from milk, with the addition of cream, buttermilk, and/or whey. This basic raw material is also called cheese milk. Cheese consists mainly of casein (the major protein of milk), other milk proteins, fat, other milk components, and a certain percentage of water, which is bound. Proteins are precipitated by enzymes and/or acidification as well as heat and are separated by suitable processes from the aqueous phase. (whey, serum). Cheese is most often molded, pressed, and salted and has fungal and bacterial culture additions. colorants, herbs, spices, and other non-dairy ingredients are also added to the cheese. It is consumed either fresh or at different stages of ripening.

Worldwide, there are more than 2000 types of cheese, sometimes made by very different manufacturing processes. A classification can be based on several aspects and is done in different countries according to different criteria. A general classification can be made for three major groups:

a. Rennet or natural cheese: manufactured straight from milk by using proteolytic enzymes (rennet) and acid, with a more or less pronounced ripening process.

b. Fresh cheese or non-ripened Cheese: Quarg, fresh cheese, and white cheese are examples. Its manufacturing process is similar to that of rennet cheese manufacturing, but it has a high degree of acidity and is not subjected to a proteolytic ripening process.

c. Long-life cheese (processed cheese): it is more often made from rennet cheese and is textured by thermal treatment and made shelf-stable.

Chemistry of Creaming

After being drawn from animals, normal milk will form a cream layer on standing. Because of the difference in density between fat globules (920 kg m−3 at room temperature) and milk plasma (density 1030 kg m−3), the granules tend to rise. This causes creaming, which we often want to prevent and it enables milk to be separated into cream and skim milk. Creaming is much enhanced when the globules have been aggregated into flocculates, clusters, or granules.

Cream Ripening

This unit operation serves as an intensive preparation of the cream for the churning process. It is divided into two categories.

1. Physical ripening;

2. biochemical ripening The following criteria are influenced by the cream ripening:

• Consistency, firmness, and spreadability of butter.

• Basic water content of butter during the manufacturing process (bound water in butter, which cannot be removed by any mechanical treatment).

• Buttermilk fat content, which has to be minimized.

The melting and solidification behavior of fat depends on the attached fatty acid, which indicate the hardness of the butterfat.

Natural hardness is caused mainly by seasonal feeding of the milk animals. Winter feed with a great deal of dry feed or roughage yields a hard butterfat with a high percentage of saturated and short-chain fatty acids and few double bonds in the fat molecules.

Summer feeding (green grass) results in fat with longer chains and a higher percentage of unsaturated fatty acids with many double bonds. We therefore distinguish between summer fat and winter fat or summer cream and winter cream.

Types of Enazymes in milk

1. Endogenous and 2. Exogenous.

Three sources of enzyme in milk:

1. The blood via defective mammary cells.

2. Secretory cell cytoplasm. Some of which are occasionally entrapped with fat globules by the encircling MFGM.

3. The outer layer of MFGM is probably the principal source of endogenous enzymes in milk. The outer layer of MFGM is derived from the apical membrane of the secretory cell which in turn originates from the Golgi membrane and acts as the principal source of enzyme in milk.

General Characteristics of Milk Enzymes

1. Deteriorative character e.g. lipase (commercially most important enzyme in milk), proteinase, acid phosphatase, xanthine oxidase.

2. Preservative enzymes. e.g. superoxide dismutase, sulfhydryl oxidase.

3. As an indicator of the thermal treatment history of milk. Related enzymes are alkaline phosphatase, ϒ glutamyl transpeptidase, and lactoperoxidase.

4. Mastitis infection indicator. Related enzymes are catalase, n-Acetyl-β D-glucosaminidase, and acid phosphatase.

5. Antimicrobial activity.

6. Commercial source of ribonuclease and lactoperoxidase

Common Milk Enzymes and their Role in Dairy Industry

The salts of the milk include those constituents that are present as ions or in equilibrium with ions. The main salts of milk are phosphates, citrates, chlorides, sulfates, carbonates, and bicarbonates of sodium, potassium, calcium, and magnesium. Approximately 20 other elements are found in milk in trace quantities including copper, iron, lead, boron, manganese, zinc, iodine, etc. These ions are more or less associated with themselves and with proteins. Some metals act as catalysts in the oxidation of milk fat leading to oxidative rancidity and certain metals like Ca, P, Mg, Mo, Co, Fe, etc. are components of metalloproteins such as lactoferrin, lactoperoxidase, xanthine oxidase, and parts of phospholipids.

Distribution of Salts Between Casein Micelles and Serum

The phosphorus is present as orthophosphate, but part of it is bound to organic components like serine and threonine residues of casein, molecules of hexoses and glycerol, phospholipids, etc. Phosphorus is also found in the hydroxyapatite structure of the colloidal calcium phosphate in the micellar structure of casein. The sulfur content of milk is about 0.36 g/kg, but most of it is in the amino acid residues methionine and cysteine of the proteins. About 10% is present as inorganic sulfate.

Physical Equilibria Among Salts

Milk contains several elements but all of them are not entirely in a soluble state. Some minerals exist in colloidal and ionic states at the normal pH of milk. The compounds that are in soluble condition play an important role in keeping various milk constituents in stable condition. A balance exists between the components that are in a soluble state and those which are in colloidal state.

Milk lipids or milk fats exist as an emulsion of small spherical droplets called milk fat globules. These globules are very small in size but very large in number. The diameter of the milk fat globules ranges between 0.1 and 22 µm and the number varies between 1.5 and 3.0 billion globules/mL. The average size of milk fat globules ranges from 2 to 5 µm. The average fat globule size in cow and buffalo milk are 3.4 and 4.5 µm, respectively. The surface of the milk fat globules is coated with an adsorbed layer of material known as the milk fat globule membrane (MFGM). MFGM contains phospholipids and proteins in a complex form. MFGM has two main functions (a) fat globule stabilization and (b) preservation of the fat globule’s identity.

General Composition

Triacylglycerols (triglycerides) represent 97–98% of the total lipids in the milk of most species. The diglycerides probably represent incompletely synthesized lipids in most cases. Although phospholipids represent less than 1% of total lipids, they play an important role in being present mainly in the MFGM and other membranous material in milk. The principal phospholipids are phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin. Trace amounts of other polar lipids, including ceramides, cerebrosides, and gangliosides, are also present. Phospholipids represent a considerable percentage of the total lipid of buttermilk and skim milk reflecting the presence of proportionately larger amounts of membrane material in these products.

Carbohydrates are polyhydroxy derivatives of aldehydes or ketones and undergo all the organic reactions common to the carbonyl groups and hydroxyl groups. The principal milk carbohydrate is lactose. It is a disaccharide. Hydrolysis of lactose with enzyme β-galactosidase yields glucose and galactose.

Lactose is the major carbohydrate present in all types of milk. The first record of isolation of lactose was in 1663, by Bartolettus by evaporation of whey. Milk contains only trace amounts of other sugars, including glucose, fructose, glucosamine, galactosamine, neuraminic acid, and neutral and acidic oligosaccharides.

Lactose plays a significant role in milk and milk products that are

• It plays a major role in the production of fermented dairy products.

• It contributes to the nutritive value of milk and milk products

• It affects the texture of certain concentrated and frozen products;

• It is involved in heat-induced changes in the color and flavor of highly heated milk products.

Status of Lactose

Lactose content varies extensively in the milk of different species. Breed of animal, individuality factors, udder infection, and especially the stage of lactation regulates the lactose content in milk. The lactose concentration in milk decreases gradually and significantly during lactation; this behavior contrasts with the lactational trends for lipids and proteins, which, after a decrease during early lactation, increase significantly during the second half of lactation. Lactose synthesis is hampered during mastitis. Lactose, along with sodium, potassium, and chloride ions, plays the principal role in controlling the osmotic pressure of the mammary system. Thus, any change in lactose content is compensated by a change in the soluble salt constituents. The osmotic relationship between lactose and soluble salts during the secretion of milk partly explains the presence of high lactose in milk with a reduced ash content and vice versa (Table 1).

Bovine milk contains an average of 3.5% protein. The concentration of protein changes significantly during lactation. The first few days of lactation exhibit the main change in protein concentration. The main changes occur in whey protein fraction.

Distribution of Protein in Milk

80% casein and 20% whey protein (β lactoglobulin- 10%, α- lactalbumin- 5%, serum albumin, and immunoglobulin- 5%).

Fractionation of Milk Proteins Fractionation of milk protein is mainly done by acidification at pH 4.6 around 30 °C.

Almost 80% of the bovine milk protein precipitates at pH4.6, and this fraction yielded by precipitation is called casein.

The remaining soluble part of the protein is referred to as whey protein/serum protein/ non- casein protein. Wide variation is observed in the casein- whey protein ratio in different types of milk(protein synthesis is independent of animal diet)

Significant Difference Between Casein and Whey Protein

1. Casein precipitates at pH 4.6 but does not precipitate whey protein at that pH. This property of milk protein is commercially exploited for the preparation of industrial casein and certain varieties of cheese.

2. Chymosin or other proteolytic enzymes produce specific changes in casein resulting in coagulation in the presence of calcium. Whey protein does not undergo this type of coagulation.

3. Casein is very stable to high temperatures. They can withstand heating up to 140 °C for 20 minutes but whey protein is relatively heat labile. They are completely denatured by heating at 90 °C for 10 min.

4. Casein is a phosphoprotein containing an average of 0.85% phosphorous while whey protein does not contain any phosphorous. The phosphate of casein is an important contributor to remarkably high heat stability and calcium- induced coagulation of rennet- altered casein.

5. Casein contains very low sulfur (0.8%) whereas whey protein is relatively rich in sulfur (1%). The principle casein contains only methionine but whey protein contains a significant amount of cystine, cysteine, and methionine. These sulfur- containing amino acids are responsible for many changes in milk on heating.

6. Casein is synthesized only in the mammary gland and no other sources in nature. Some of the whey proteins β lactoglobulin, and α- lactalbumin are synthesized in the mammary gland but bovine serum albumin and immunoglobulin are not synthesized in the mammary gland but derived from blood.

7. The whey protein is dispersed in solution and has a simple quaternary structure. Casein has a complex quaternary structure and exists in milk as large colloidal aggregates.

Introduction

The mammary glands of female mammals secrete milk, a necessary food that is liquid and meets the newborn’s nutritional needs. All mammals consider milk to be a complete food for their newborns. It is considered a necessary food for humans and can be consumed as a product or as liquid milk. It contains almost all essential nutrients except iron, which is very low in milk. In essence, milk is an emulsion of protein and fat in water, supplemented with carbohydrates, vitamins, and minerals.

Definition of Milk

• Milk is a complex biological fluid that differs from species to species in both composition and physical properties. It is the full lacteal secretion of a mammary gland that is obtained by milking mammals for a minimum of 72 h after calving or until the milk is free from colostrum.

• FSSAI defines milk as the normal mammary secretion derived from the complete milking of healthy milch animals. It should be free from colostrum.

From a chemical perspective, milk is a dynamically balanced mixture that comes in different forms such as emulsion, colloidal suspension, and true solution. It also contains varying amounts of proteins, fats, carbohydrates, salts, and water.

Gross Composition of Milk

Milk composition varies among different mammals. The average chemical composition of milk in various species is given in Table 1. Buffalo milk and sheep milk contain higher amounts of fat, while human milk comparatively has the highest lactose content. Human milk has lower protein content as compared to milk of other species.

A laboratory is essential for research work in science and development. Most of the research institutes have dedicated laboratories for researchers and students to conduct experiments, perform research work, as well as to provide hands-on learning/training to students. There are certain rules and regulations laid down by the laboratory in-charge which need to be followed strictly for safety purposes. Some of the laboratories keep ‘safety manual’ for researchers to read before entering into the laboratory to conduct experiments. Whenever safety is compromised in a laboratory, there is increase in risk of unwanted incidents in the laboratory. Storing food/beverages in the same refrigerator where laboratory chemicals are kept may lead to disastrous effects on health. This chapter aims to provide instructions in brief on the things to be performed (Do’s) before and after entering onto a laboratory as well as the things never to be done (Don’ts) in a laboratory for the safety reasons.

Good Laboratory Practices (Do’s)

• P Wear apron/labcoat while working in the laboratory.
• P Use gloves/respiratory mask/fumehood while handling toxic/volatile/inflammable chemicals/reagents.
• P Use pippet or pippetor for safety in transfer of solutions.
• P Protect eyes, face and naked parts of the body from UV radiations. Use safety goggles or face mask/apron.
• P Keep your eyes away from Liquid Nitrogen and use cryogloves while handling it.
• P Arrange all the necessary materials/solutions before starting the experiment.
• P Periodically fumigate/ozonize culture and inoculation rooms.
• P Autoclave all contaminated materials before discarding.
• P Pre-treat gel or solution containing Ethidium Bromide (EtBr) before discarding into laboratory waste-disposal bin, and use gloves while handling them.
• P Burn all the transgenic plant materials in an incinerator after taking necessary observations.
• P Make entry in the designated log-book and clean the instrument after using it properly.
• P Switch-off the instrument after use or when not in use.
• P Wash hands with soap and remove the labcoat before leaving/going out of the laboratory7.
• P Ensure that switches for fans/lights, and water taps are off while leaving the laboratory.
• P Follow the biosafety guidelines suggested for handling and disposal of the hazardous chemical.

Introduction

Several hazardous chemicals, microbes, and techniques like recombinant DNA technology are used for research and development work in research Institutes and universities. Therefore, the researchers, technical, helping and supporting staffs as well as the laboratory outputs must be kept free from contaminations. Thus, biosafety is necessary to prevent unintentional exposure/ release of hazardous chemicals, microbes/pathogens, and toxins to the laboratory staff and environment. Following the biosafety guidelines and ethical considerations are essential to maintain safety standards for the researchers as well as the environment. This chapter deals with various biosafety measures and ethical issues to be strictly followed while working in a research laboratory. The required safety level needs to be determined as per the chemical/biological materials are used in the laboratory to keep the researcher/environment safe for sustainable research and developmental activities.

Biosafety

The term biosafety is used to describe the procedure and policies needed to ensure environmental and personal safety. Biosafety refers to the containment principles, technologies and practices that are implemented to prevent unintentional exposure to pathogens, toxins, or their accidental release into the environment. A fundamental objective of any biosafety program is the containment of potentially harmful chemical/biological agents as precautionary safety measures.

Why Biosafety?

With the increasing number of countries adopting molecular biology tools and techniques in life science research and development activities, especially in the areas of agriculture and medicine, the biosafety issues are gaining importance to ensure biological safety for the public and the environment. Conducting research in safe and sustainable manner is not only a personal requirement but essential collective efforts to ensure biological safety for a clean and safe environment. This requires rules, regulations, monitoring bodies, and awareness among the workers as well as the public. Therefore, biosafety per se is an integral part of laboratory research and requires awareness among the researchers so that biological safety can be well taken care from the grassroot level.

For a researcher, laboratory is another home where he/she spends much time every day. As we take care of cleanliness, safety, and design of our home, we must take care on these aspects of laboratory too. It is the responsibility of the laboratory personnel to ensure cleanliness/safety of the laboratory. Since every laboratory chemical and equipment have the potential to cause harms if not used with appropriate precautions. Hence, as a laboratory user, it is our responsibility to follow the basic safety/biosafety rules and regulations set up by the Principal Investigator of the laboratory, Institutional Biosafety Committee, as well as by other regulatory authorities, if any. We must be aware of the general/personal protective measures, cleanliness and laboratory waste disposal prior to joining a research laboratory. In this chapter we will briefly discuss about personal safety, chemical safety, cleanliness, waste disposal, etc.

A. Personal Safety

1. Personal Protective Equipment (PPE)

Personal protective equipment (PPE) is special arrangement to protect the worker from specific hazards because of using a hazardous substance. PPE does not reduce or eliminate the hazard, but protects the wearer from exposure to hazardous material. PPE should not be reused or brought outside the laboratory as these might increase the risk of spread hazardous material or contamination. Used PPE must be kept inside the laboratory in a designated waste storage area adjacent to the work area.

Introduction

Proper electrical wiring and earthing are crucial for laboratory safety and functionality. A laboratory houses several installations/equipment that demand stable/continuous power supply. Inappropriate or faulty wiring and earthing may lead to significant risks, including electrical shocks, damage to equipment, and even incidence of fire. Therefore, understanding the requirements of electrical wiring and earthing is essential for designing, construction, and maintenance of laboratory. This chapter aims to provide a basic knowledge of electrical wiring and earthing necessary for a laboratory. Importance of earthing in mitigating electrical hazards and ensuring integrity of electrical installations are also described.

Basics of Electrical Wiring

Basic understanding of electrical wiring is necessary for laboratory safety and efficient electrical functioning. The potential difference between two electrical points (negative and positive) is known as Voltage, which drives the flow of electric current. This is measured as volt (V). The rate of flow of electricity in a circuit is known as Current (I), which is measured as ampere (A). Current can be either direct (DC) or alternating (AC). Opposition/resistance experienced in the flow of current in circuit is known as Resistance (R) and it is measured in ohms (Ω). Higher the resistance means lesser the current flows at a given voltage. The rate at which electrical energy is transferred/utilized in an electric circuit is known as Power (P), which is measured in watt (W). The Power can be calculated as the product of voltage and current (P = V × I).

Introduction

We come across a variety of chemicals every day in a laboratory. While some of them are relatively safer, others are extremely hazardous. Thus, it is very important to know the potential hazards that can be caused by the chemicals present in the laboratory and the safety measures needed to be followed to avoid the hazards. Additionally, we should also be aware of the steps necessary to be taken on such hazardous situations. According to IUPAC, a hazardous chemical mean the chemical which has the potential to cause damage in various ways. In a laboratory, there are chemicals that might be irritant, carcinogen, respiratory fibrogen, systemic poison, asphyxiant, flammable, etc. which have the potential to cause hazard to the workers or environment. Therefore, it is important to use appropriate procedure to reduce the risk and protect the health/safety of laboratory personnel, the public and the environment.

Types of Hazards Associated with Chemicals

Chemical hazards: The hazards caused by chemicals can be broadly classified into three categories.

1. Health hazard: This is also known as toxic effects of chemical. The risk associated with a chemical in laboratory depends on the extent of exposure and the inherent toxicity of the chemical. Many chemical reactions form the product that might be much more toxic than the reacting chemicals. For example, inadvertent mixing of formaldehyde (a common tissue fixative) and hydrogen chloride results in the generation of bis(chloromethyl) ether, a potent human carcinogen. For most of the chemicals, the route of exposure (through skin, eyes, gastrointestinal tract, or respiratory tract) is more important (Figure 17.1) that must be considered during risk assessment.

Introduction

Flowering plants (Angiosperms) are identified by their morphological characteristics like flower, fruit and seeds that help their taxonomic characterization into different species, genera, families and orders. The floral characters which are used for taxonomic characterization of plants include the type of inflorescences, perianth structure, floral symmetry, arrangement of floral leaves, type of androecium, stamens, gynoecium, carpel, ovule, etc. The type of fruit and seed also play important roles in taxonomic classification. In this chapter we will briefly discuss about various morphological features of flowering plants (in botanical terms) that are useful in taxonomy.

Nomenclature of Plant

Nomenclature of plant is the formal and scientific naming of plants. It is related to but distinct from taxonomy for grouping and classifying plants and provide botanical name.

The scientific/botanical nomenclature started with Species Plantarum by Carolus Linnaeus in 1753. Botanical nomenclature is governed by the International Code of Nomenclature for algae, fungi, and plants (ICN), which replaces the International Code of Botanical Nomenclature (ICBN).

ICBN and ICN

International Code of Botanical Nomenclature, (ICBN) was adopted by the VIIth International Botanical Congress held in Stockholm, July1950. While the International Code of Nomenclature for algae, fungi, and plants (ICN) was adopted by the XVIIIth International Botanical Congress held in Melbourne, Australia in July 2011.

Introduction

Test of pollen viability (extent of pollen viability) is important in pollination experiments to ensure fertility/seed setting. It is also important in monitoring state of pollens during storage in ecological/ taxonomic studies as well as during research on pollen biochemistry, genetics, stigma interactions, and incompatibility studies. Pollen viability refers to the ability of pollen to accomplish post-pollination events to achieve fertilization. Pollen viability test has gained importance particularly in plant breeding and hybrid seed production experiments/ practices. In this chapter, you will be acquainted with the basics of pollen viability test. However, for more professional knowledge on the topic readers are suggested to refer specialized books.

Pollen Viability Test

It is important to know the extent of viability of pollens used in pollination experiment, in finding fertility of a plant of interest, to monitor state of pollen on storage, in ecological/taxonomic studies, as well as in research on biochemistry, genetics, stigma interaction, incompatibility assay, etc. of pollen. Several pollen viability tests have been developed time-to-time. It is recommended to initially use different tests for a given pollen type to find out the most appropriate method that provides reliable result on pollen viability. It is important to note that every procedure requires standardization of protocol according to the sample used. Following different tests are used for assessing pollen viability.

1. In-vitro Pollen Germination Test

It is a simple, speedy, and quantitative test for assessing pollen viability. In this method, a collection/sample of pollen is germinated in vitro and observed under microscope. The percentage of germinating pollens (producing pollen tube) after appropriate treatment and a given period of time is determined. The percentage provides an index of viability of the pollen sample.

Introduction

Seed is a living entity (mature ovule) consisting of a tiny plant in embryonic stage along with the stored food protected by an outer layer known as seed coat. Seed is an important biological input in agriculture, which is economically important for successful crop husbandry. Cotyledons serve as the source of nutrients (proteins, carbohydrates, and oils) that not only nurture the plant at the early developmental (seedling) stages of plant but also the vegetarian (heterotroph) organisms. Being one of the important inputs in agriculture, viability of seed needs to be assessed to know the germination percentage of seeds used to raise a crop. Since viability of seeds may deteriorate on storage depending on the storage conditions. Therefore, often it becomes important to test viability/germinability and vigour of seeds before being used for raising a crop or conducting an experiment. In this chapter, we briefly discuss about the different methods which can be used for testing the viability of seeds.

Seed Viability

Seed viability can be defined as the capability of the seed to germinate and produce a normal seedling within a specified period. In other words, seed viability indicates how many seeds in a lot are alive and could develop into plants under appropriate field conditions. It is very important that most of the stored seeds are capable of germinating when sown in a field. They must possess higher viability at the beginning of storage and maintain it over the storage. A seed with high initial viability survives longer in storage; however, the viability deteriorates slowly at first and then rapidly as the storage prolongs. It is important to know the duration of storage after which a considerable decline in viability occurs under given storage conditions so that to take necessary action to revive the seeds. Excessive deterioration in vianility of seeds may lead to loss of the seed/material/ germplasm.