eChapters

India is bestowed with a wide variety of livestock species which forms the backbone of the economy of majority of the farmers. Cattle are one of the important species domesticated by man mainly for their draftability in agricultural operations. Rearing of cattle is an integral part of the Indian agriculture as it is always considered as a subsidiary enterprise of the agriculture farming. The indigenous cattle are well known for their adaptability to the harsh tropical climatic conditions, ability to thrive in the resource poor production systems, better feed efficiency and disease resistance. Barring few dairy breeds, majority of the Indigenous cattle breeds are poor in their genetic potential for milk production. Considering the ever increasing demand from milk , the country has implemented many cattle improvement programmes like Central Herd Registration Scheme (CHRS), Intensive Cattle Development Programme (ICDP), All India Coordinated Research Project (AICRP) on Cattle National Programme for Bovine Breeding and Dairy Development (NPBBD), National Livestock Mission, National Project for Cattle and Buffalo Breeding (NPCBB), Rashtriya Gokul Mission (RGM) etc., for improving the milk productivity of cattle.

True /False

Q.1.    A diagnostic test enables the doctors to reach at a decision whether the patient is suffering from a particular disease or not.
Q.2.    The recent developments in the field by biotechnology have led to the development of many new medical tests.
Q.3.    The general principles in designing and conduct diagnostic test studies are not known well.

True/False

Q.1.    Demographic data and vital statistics are important tools for researchers, epidemiologists, health planners and health professionals.
Q.2.    The distribution of people across different geographic regions is known as population density.

The term “artificial intelligence” (AI) refers to a wide variety of theories and technologies that are used to the resolution of issues characterized by a high level of logical or algorithmic complexity. It cuts across a wide range of f ields, such as mechanical modeling, software engineering, data science, and statistics. Many artificial intelligence (AI) technologies, which first appeared in the 1950s, have lately been refined or extended thanks to advances in computer performance. Recent advancements have been made possible by the creation of interfaces between artificial intelligence and other fields of study, such as biomedicine, and by the collection of enormous amounts of data from a variety of fields, notably those related with the healthcare industry. First, Artificial intelligence helps with situational awareness, which is relevant in animal health and animal husbandry because it allows for the detection of patterns like repeated sequences of observations, forms like a protein, and signals like increased mortality especially in comparison to a baseline at various scales. A third option is computer-based decision making, or more practically, human decision support via knowledge - based systems, diagnostic support, and resource allocation, for instance.

Significant advancement in high throughput technologies (microarrays, automated sequencing and mass spectrometry), transcriptomics, the global study of transcription, together with genomics and proteomics, has undoubtedly contributed to a systems biology approach. These technologies have generated vast amount of sequence data from plants, animals and microorganisms. Fortunately, efficient computational tools (intelligent data networks, query, retrieval, analysis and visualization tools) have now optimized data mining, accelerating the process of discovery. The analysis of sequence variation is of major importance in genetic studies. In this context, molecular markers represent one of the most powerful tools for the analysis of variations in the genome and the association of heritable traits with underlying genetic variation. The development of high-throughput methods for the detection of simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs) has led to a revolution in their use as molecular markers. The availability of large sequence data sets permits mining for these molecular markers, which may then be used for numerous applications such as genetic mapping, association mapping, evolutionary and phylogenic studies.

Introduction

The branch of science that deals with resolving the evolutionary relationships among organisms is phylogenetics. Phylogeny is the description of biological relationships, usually expressed as a tree. A statement of phylogeny among objects assumes homology and depends on classification. Phylogeny states a topology of the relationships based on classification according to similarity of one or more sets of characters, or on a model of evolutionary processes. In many cases, phylogenetic relationships based on different characters are consistent, and support one another. If different characters induce inconsistent phylogenetic relationships, they all are dubious. Conversely, note that the same similarity data may be consistent with different possible topologies or trees.
 
Molecular approaches to phylogeny developed against a background of traditional taxonomy, based on a variety of morphological characters, embryology, and from fossils, information about the geological context (stratigraphy).

Introduction

Natural evolution gifted the biological world a treasure of diverse bio-molecules that take part in various cellular activities. Proteins, one of the vital molecules of biological function are wisely folded polymers comprising 20 different aminoacids. Function of each protein is encoded in its aminoacid sequence. Aminoacid residues can indulge in bonded or non-bonded interactions depending on the environment. These inter residue interactions with in a polypeptide drive its folding in to a compact conformation from a linear molecule. Biochemical properties as well as the linear arrangement of the aminoacids determines the three dimensional structure of a protein. These determinants of defined structure directly or indirectly dictates functional fate of the folded protein also.

I. MANDATE OF COPYRIGHT LAW

The importance of intellectual property rights has arisen in recent years to a great extent. The law of copyrights protects original forms of expression i.e. Novels, Movies, Musical compositions and computer software etc. Earlier it was mainly confined to the -published Work and owner of it could prevent others from getting it published in the form of books or articles in order to get benefit in terms of money or goodwill or reputation. In case of such undue enrichment, the owner had a valid cause of action. The owner of the copyright has a valid cause of action against the violator. At present even in case of its adoption and translation or abridgement, it is obligatory to have consent of author. The main purpose and object of copyright law is to protect the expression in any other form from unauthorized use. Since 1847, the copyrights law is in developing stage so, many legislations has been enacted.1 With the changing of times, the meaning of expression also changes and the style of exhibit the things also changed. The right of author who gives expression in any form is protected from unauthorized use.

5.1 Plant Breeder’s Rights (PBR) and Protection of Plant Varieties and Farmers’ Rights (PPV&FR) Act
5.1.1 Introduction
In the context of biotechnology and agricultural innovation, Plant Breeder’s Rights (PBRs) offer exclusive intellectual property protection to individuals or organizations that breed, develop, or discover new plant varieties. These rights ensure economic incentives for plant breeders while promoting innovation in seed development and agricultural productivity. In India, the unique and inclusive Protection of Plant Varieties and Farmers’ Rights Act (PPV&FR), 2001, balances breeders’ rights with recognition of farmers’ contributions, making it one of the most progressive legislations in the world.
5.1.2 Concept of Plant Breeder’s Rights (PBR)
Plant Breeder’s Rights refer to the exclusive rights granted to plant breeders to produce, sell, market, distribute, import or export their registered plant varieties. These rights are sui generis, meaning they are a special form of IP distinct from patents, developed to cater specifically to plant varieties.
Key Objectives of PBR
• Encourage investment in plant breeding and biotechnology.
• Ensure access to improved crop varieties.
• Prevent unauthorized commercial exploitation of novel varieties.
• Facilitate biodiversity conservation through formal registration systems.

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.

Introduction
It is a fundamental biological process in which one cell splits into two or more daughter cells. Both unicellular and multicellular organisms rely heavily on it for growth, development, repair, and reproduction. There are two main types of cell division: mitosis and meiosis. Mitosis occurs in somatic cells and produces two genetically identical daughter cells that share the same chromosomal number as the parent cell. This process is necessary for tissue development and repair. Meiosis, on the other hand, happens in germ cells and results in the formation of gametes, which reduces the number of chromosomes by half to ensure genetic diversity in progeny. Proper regulation of cell division is vital for maintaining cellular health, and disruptions in this process can lead to disorders like cancer.
The different stages of cell division is as follows:-
1. Interphase
• Although not a component of mitosis, interphase is essential for preparing the cell for division. It is separated into three sub-phases:
• G1 phase (Gap 1): The cell expands in size while creating proteins and organelles. S phase (Synthesis): DNA replication occurs, guaranteeing that each daughter cell receives an identical copy of the genetic information.
• G2 phase (Gap 2): The cell continues to expand while new organelles are synthesised)
The cell checks for DNA replication mistakes and gets ready for mitosis.
2. Prophase
• In this phase chromatin condenses into visible chromosomes, each made up of two sister chromatids linked at the centromere.
• The nuclear membrane breaks down, allowing the spindle fibres to interact with the chromosomes.

Introduction
Gametogenesis is the process by which gametes are produced in sexually reproducing organisms. This process involves a series of cellular divisions and differentiations and can be divided into two main types: spermatogenesis (the production of sperm) and oogenesis (the production of ova). Gametogenesis ensures genetic diversity through the processes of crossing over and independent assortment during meiosis. A gamete is a specialized cell involved in sexual reproduction. Gametes are haploid cells; contain only half the number of chromosomes of a diploid cell. Two gametes one from each parent fuses during fertilization to form a zygote, a diploid cell with a full set of chromosomes. The primordial germ cells develop from the wall yolk sac and differentiate into female and male gametes. During and after migration of primordial germ cells, they proliferate by mitoses and male and female primordial germ cells surrounded by somatic cells of their specific gonads and differentiate into oogonia in ovary and spermatogonia in testis. Meiosis is the process of cell division (Meiosis I and Meiosis II) in two specialized germ cells i.e. oogonia and spermatogonia to produce haploid female gametes (ovum) and male gametes (sperm) respectively. Meiosis I allow genetic recombination where exchange of DNA segment occurs between maternal and paternal chromatids.
Spermatogenesis
The development of sperm in the germinal epithelium of seminiferous tubules of the testis is known as spermatogenesis. Spermatogenesis starts, when the male animal attains the puberty. The sequential stages in spermatogenesis are
1. Mitotic/multiplication phase: Primordial germ cells undergo a series of mitotic divisions and localized in solid cords of primitive sustentacular cells (progenitor of Sertoli cells) within the seminiferous tubules (testis). At the age of puberty, the cell cords differentiated into seminiferous tubules of the testis and simultaneously primitive sustentacular cells became Sertoli cells and primordial germ cells produce spermatogonia throughout the life, having the features of stem cells. These spermatogonia remains dormant until the onset of puberty. Sertoli cells are very important for both physical support and paracrine regulation of spermatogenesis, and also form the blood-testis barrier by sealing off the seminiferous tubules with tight junctions. At puberty, they are activated and undergo series of mitotic division and produce clones of cells referred to as type A spermatogonia. Among type a

Introduction
The mammalian embryos are subdivided into two groups namely domestic animals where extraembryonic membranes form by folding process, and primates where blastocyst implants very soon after it enters the uterus and extra embryonic membranes forms after implantations. The major function of fetal membranes is to provide nutrients to developing fetus, protection, synthesis of enzymes and hormones and caring of fetal waste products. It is consisted of yolk sac, amnion, allantoic and trophoblast or chorion. The trophoblast is the single most important tissue of fetal membranes which has potential role in absorbing, transmitting and handling nutritive and waste products.
Yolk sac
It is a primitive structure. Develops early in the embryonic period from the entoderm which disappears after short period of time in ruminants. The uterine milk, secreted by the endometrial glands, is absorbed by the blastocyst along with yolk sac to provide nutrition to early embryo. Before forming other embryonic membranes yolk sac performs limited functions whose role is limited during mid and late gestation.
Amnion
It is an ectodermic vesicle that arises from an out folding of the chorion which is a double walled sac that completely surrounds the fetus except at the umbilical ring. The inner layer of this double-walled sac is the “true amnion” and the outer layer is the “false amnion” amnion chorion. Like yolk sac, the amnion chorion also lasts for shorter period.The amniotic fluid is clear, colorless and mucous in nature. Source of amniotic fluid: In early to mid-gestation, it is probably from the amniotic epithelium and from fetal urine as the fluid is quite watery. As gestation advances, the allantoic fluid increase in volume while the amniotic fluid volume remains fairly static but

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.

11.1 Principles and Framework for Risk Assessment
11.1.1 Introduction
Risk assessment is an indispensable scientific tool in evaluating the safety and potential impacts of genetically modified (GM) crops on human health, animal welfare, and the environment. It provides a structured, evidence-based framework for identifying hazards, characterizing risks, and proposing mitigation strategies before the approval and commercialization of GMOs. In the context of biotechnology and regulatory biosafety, risk assessment ensures that GM crops meet both national and international standards for biosafety and sustainability.
11.1.2 Definition of Risk Assessment
Risk assessment is defined as the systematic process of identifying, evaluating, and characterizing the potential adverse effects arising from exposure to a GMO or its products, under specified conditions of use.
Risk = Hazard × Exposure
Where hazard is the potential to cause harm, and exposure is the likelihood of encountering the hazard.
11.1.3 Objectives of Risk Assessment in GM Crops
• To evaluate the safety of GM crops for human and animal consumption.
• To assess the potential environmental impacts, including effects on biodiversity, gene flow, and ecosystem interactions.
• To ensure that regulatory decisions are science-based, transparent, and precautionary.

Cell culture medium : To grow cells in in vitro, environment should be as close as possible to in vivo condition. But a number of factors are important to grow cells in vitro namely substrate, temperature and medium. There is a number of ready made medium available commercially and made the cell culture work easier. Any successful medium consists of isotonic buffer solution containing inorganic salts, amino acids, vitamins, a energy source and various supplements. Usually serum or serum replacement is used as supplement.

Introduction
• Overview of the development of the musculoskeletal system, including bones, cartilage, joints, and muscle types within skeletal and cardiac systems.
Development of the Musculoskeletal System
Mesoderm Contributions
• Mesodermal Derivatives
• Gives rise to skeletal muscle, skin dermis, endochondral bones, and joints.
• Notochord and Somite Formation
• The notochord induces the paraxial mesoderm to form somites, with somitomeres developing rostrally to the notochord in the head)
• Somite Differentiation
• Each somite differentiates into three regions:
• Sclerotome (medial): Forms most of the axial skeleton (vertebrae, ribs, base of the skull).
• Dermatome (lateral): Migrates to form the dermis of the skin.
• Myotome (middle): Migrates to form skeletal muscles. Individual adult muscles arise from the merger of adjacent myotomes.
Muscle Development
• Origins of Muscle Tissue:
• All skeletal muscle derives from paraxial mesoderm forming somites and somitomeres, with the exception of iris musculature (derived from optic cup ectoderm).
• Cardiac and smooth muscles originate from splanchnic mesoderm.

The difficulty in conducting a factorial experiment in an RBD is that, as the number of factors and/ or that of levels of the factors increase, the number of treatment combinations to be compared increases too. This in turn necessitates use of large sized blocks to accommodate all the treatment combinations i.e in a 2^4 – experiment there should be 16 plots in a block. But it has been found that the experimental error increases with an increase in the size of a block, for then it becomes less effective in controlling the heterogeneity of the units. A remedy has been found out : this is to divide a replicate (a complete block) into a number of equal blocks (incomplete blocks) and then to allocate the treatment combinations to these blocks so that only the unimportant treatment comparisons gets mixed up or entangled with the block comparisons. These treatment comparisons are then said to be confounded or mixed up with block effects. These effects cannot be separately tested or estimated. But the remaining treatment effects, which are not confounded with the block effects, are still capable of separate estimation and testing.

15.1 Biopiracy – Definition, Examples, and Impacts
Introduction
Biopiracy represents a grave ethical and legal concern in the field of biotechnology and intellectual property rights. It refers to the unauthorized and uncompensated exploitation of biological resources and traditional knowledge—primarily from developing countries—by corporations, research institutions, or individuals, often for commercial gains. Biopiracy is inherently unjust as it involves the misappropriation of community knowledge, undermining the rights of indigenous peoples, and violating the principles of equity, fairness, and sovereignty over natural resources.
As globalization intensifies biotechnological exploration, biopiracy challenges ethical norms and highlights the need for strong legal, institutional, and ethical safeguards.
Definition of Biopiracy
Biopiracy can be defined as:
“The unauthorized extraction, use, and commercialization of genetic resources or associated traditional knowledge from indigenous communities or biodiversity-rich regions without prior informed consent or fair and equitable benefit sharing.”
Biopiracy often involves patenting biological materials or indigenous medicinal knowledge without recognizing the origin communities as rightful holders of such knowledge or genetic wealth. It is closely linked to ethical misappropriation, exploitation, and violations of bioethical norms and international agreements.

1.1     Prokaryotic 
    
1.2     Eukaryotic
    
1.3     Cell orgenellie