eChapters

This chapter examines immune-system failure arising from reactions against self-antigens or inadequate immune responses. It introduces immunological tolerance, central and peripheral tolerance, clonal deletion and clonal anergy. Important autoimmune diseases—including myasthenia gravis, Graves’ disease, systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis and Sjögren’s syndrome—are considered. The chapter also discusses molecular mimicry and the production of autoantibodies. Primary, secondary and severe combined immunodeficiency disorders are differentiated, with examples such as Chédiak–Higashi syndrome. Immune-privileged sites and immunological paralysis are also covered. These topics explain the mechanisms that normally prevent self-reactivity and the consequences that arise when immune regulation becomes defective.

Introduction

Avian medicine is a rapidly growing field, with an increasing number of exotic bird species being kept as pets. However, the unique anatomy, physiology, and behaviour of these birds present several challenges as well as new opportunities for veterinarians and researchers.Now a day’s birds are gaining popularity over dogs and cats because of various reasons listed below:

a. They are more suited in present life style as can be suited well in small spaces.

b. They require low maintenance with relative low cost of maintenance and live longer life.

c. They form very strong bonds with their owners.

Birds can be classified into different families, such as Psittaciformes, Passeriformes, Columbiformes, Piciformes, Anseriformes, Galliformes etc. containing numerous species (>10,000). The first documentation of parrot as a pet was found in Rig Veda, written more than 3000 years ago. Evidence of bird keeping is observed in ancient civilizations of Greece, Rome, China and Egypt. In Greek mythology, Raven (a crow) was mentioned as a symbol of good luck and was associated with God Apollo (God of prophecy). Crows are also considered as ancestors in Hindu mythology and food (pinda) is offered to crows during ritual of paying homage to dead persons. In most of the countries psittacine and passerine birds are preferred as pet birds.

Introduction Antimicrobial resistance (AMR) has emerged as one of the gravest threats to modern medicine, undermining decades of progress in infectious disease control. It is cited by the World Health Organization (WHO) as one of the top 10 threats to global health. In 2019 alone, AMR was associated with approximately 4.95 million deaths, including 1.27 million directly attributable to resistant bacterial infections. This makes AMR a leading cause of death worldwide, surpassing HIV/AIDS (0.86 million deaths) and malaria (0.64 million deaths) in the same year. Projections suggest that if the trend continues unchecked, AMR could cause 10 million deaths annually by 2050, representing a profound human-cost escalation. Livestock and Animal burden While the quantitative burden of AMR in animals is not yet globally mapped, its effects in livestock are already both profound and alarming. Over 70% of all antimicrobials globally are used in farm animalsfrequently for growth promotion and disease prevention—not just treatment. This widespread use contributes to emerging resistance in key pathogens—Salmonella, E. coli, Campylobacter in poultry, and severe swine dysentery in pigs. In some outbreaks, swine dysentery, resistant to conventional treatment, has caused up to 90% morbidity and 30% mortality in weaned pigs. Animal-to-Human Transmission and the One Health Paradigm Antimicrobial resistance in livestock doesn’t stay on the farm—it seeps into the environment and crosses over to humans via food chains, direct contact, and environmental pathways. For example, substandard veterinary medicines and antibiotic misuse has been traced in zoonotic infections and veterinary-tohuman AMR transmission.Surveillance gaps remain: the World Organisation for Animal Health (WOAH) acknowledges that while human-related AMR mortality is well-documented, the animal health burden is not yet quantified globally—though numerous projects are underway to address this gap. The failure of conventional antibiotics to treat resistant infections highlights the urgent need for alternative therapeutic strategies. Among the most promising options is bacteriophage therapy—the use of viruses that specifically target and kill bacteria. Bacteriophages Bacteriophages, often referred to simply as phages, are viruses that infect and replicate within the bacteria. They are considered as the most abundant biological entities on Earth, with an estimated amount 10³¹ phage particles present in the biosphere—outnumbering bacteria by approximately tenfold. Phages are naturally found in diverse ecosystems including soil, ocean, sewage, and even within the microbiomes of humans and animals.

Accuracy

The degree of closeness of the determined value to the nominal or known true value under prescribed conditions. This is sometimes termed trueness.

Analyte

A specific chemical moiety being measured, which can be intact drug, bio molecule or its derivative, metabolite, and/or degradation product in a biologic matrix.

Analytical run (or batch)

A complete set of analytical and study samples with appropriate number of standards and QCs for their validation. Several runs (or batches) may be completed in one day, or one run (or batch) may take several days to complete.

Biological matrix

A discrete material of biological origin that can be sampled and processed in a reproducible manner. Examples are blood, serum, plasma, urine, faces, saliva, sputum, and various discrete tissues.

Flow Cytometry is a tool used to detect and measure chemicophysical characteristics of a population of cellular particles. Modern day biomedical research needs the molecular assembly of tools to proceed in the advent of biomedical research with real time diagnostic, prognostic and experimental detection of markers. For the same, a naïve tool is flow cytometry. In this technique, a sample having cells or particles is suspended into a fluid and injected into instrument. The ability to perform these measurements in a very rapid time span is one of the key advantages of the flow cytometry process. A single-cell suspension must first be prepared in order to analyse solid Tissues. A flow cytometer is look alike to microscope, except that, instead of producing an image of the cell, it generally offers a cell-by-cell basis high-throughput, automated quantification of specified optical parameters.

They can quantify up to three to six properties or components are quantitated in a single sample, cell by cell, for about 10,000 cells, in less than one minute. Sample is focused to ideally flow one cell at a time through a laser beam, where the light scattered is characteristic to the cells and their components. These cells are tagged with fluorescent markers in order to absorb the light and then emitted them in the form of band of wavelengths. Average ten thousand cells can be quickly examined and the data gathered are processed by a computer.

During recent years ultrasonography has become an indispensable diagnostic modality in human and veterinary medicine because of its non-invasive nature, providing visualization of internal organs, and quick results. Birth of ultrasound took place in 1794 when an Italian biologist Lazzaro Spallanzani discovered that bats used sound waves for navigation. Since then continuous developments and advancements in the technology have made the sonography a dependable and an easily accessible diagnostic modality.

This chapter explains how B-cell receptors, T-cell receptors and major histocompatibility complex molecules participate in antigen recognition. It differentiates MHC class I and class II pathways and describes the processing of endogenous and exogenous antigens. The roles of dendritic cells, macrophages and B lymphocytes as professional antigen-presenting cells are examined. Topics include MHC restriction, antigenic peptides, proteasomal degradation, ubiquitination and recognition by CD4-positive and CD8-positive T lymphocytes. The chapter also discusses superantigens, opsonization, phagocytosis, T-independent antigens and species-specific MHC systems such as BoLA and the avian B complex. These concepts explain the initiation of adaptive immune responses.

1. Introduction

Malaria remains one of the most serious tropical diseases and kills an estimated one to two million people each year worldwide (Snow et al., 2005). Spread of drug resistance has played an important role in the occurrence and severity of epidemic diseases in the world (Majori, 2004). As a consequence of resistance to drugs such as quinine, chloroquine, primaquine and mefloquine, the vehemence of malaria has increased in many endemic regions of the world (Schlitzer, 2007; Basco et al., 1994). One of the biggest challenges to malaria control is the emergence and spread of drug resistant P. falciparum strains. Although human malaria transmitted by female Anopheles mosquitoes has four Plasmodium species as its aetiological agents P. falciparum, P. vivax, P. ovaleand P. malariae, the most widespread and severe disease is caused by P. falciparum, which transiently infects the liver before invading red blood cells of the mammalian host. Clinical manifestations occur at the erythrocytic stage and can include fever, chills, prostration and anaemia, as well as delirium, metabolic acidosis, cerebral malaria and multi-organ system failure, which may be followed by coma and death (Fidock et al., 2004; Deprez-Poulain et al., 2005, De Sa et al., 2009).The success of the natural product-based antimalarials such as quinine and artemisinin has showed the importance of plants as potential source of antimalarial agents. Consequently, numerous efforts to identify antiplasmodial activities from plants traditionally used as remedies for fever and malaria (Chen et al., 1997; Havsteen, 2002;

1. Introduction 
Among the innumerable gifts of nature, plants, animals and minerals have been the basis of treatment of human diseases. Generally it is either of prebiotic origin or originating from microbes, plants or animal sources. More likely they are the natural expression of the increase in complexity of organisms. Beside animals, plants are one of the most important sources of natural products. From the very beginning of their existence, man has acquainted himself with plants and used them in a variety of ways throughout the ages. In search of food and to cope successfully with human sufferings, ancient man began to distinguish those plants suitable for nutritional purposes from others with definitive therapeutic action. Some plants are widely used as food while others showed beneficial effects against various human sufferings such as injuries and diseases. This relationship has grown between plants and man, and many plants have come to be used as drugs. The use of plant as a drug can be divided broadly into five periods. The early period covers the Indian, Chinese, Sumerian, Egyptian and Assyrian civilizations followed by the Greco-Roman, Arabian, Medieval and modern periods (Buchnman, 1980). Among ancient civilisations, India has been known to be rich repository of medicinal plants as the forest in India is the principal repository of large number of medicinal and aromatic plants, which are largely collected as raw materials for manufacture of drugs and perfumery products. About 8,000 herbal remedies have been codified in Ayurveda. The Rigveda (5000 BC) has recorded 67 medicinal plants (Chopra et al., 1956), Yajurveda 81 species and Atharvaveda (4500-2500 BC) 290 species. Charak Samhita (700 BC) and Sushrut Samhita (200 BC) had described properties and uses of 1100 and 1270 species, respectively (Kirtikar & Basu, 1918; Baquar, 1989) in compounding of drugs and these are still used in the classical formulations, in the Ayurvedic system of medicine. Before the availability of synthetic drugs, medicinal plants were used as the most primary source of medicine and continue to provide the mankind with new remedies for many diseases. These were used in crude forms like expressed juice, powder, decoction or infusion. As the plant have specialized biochemical capabilities they are able to synthesize and accumulate a vast array of primary and secondary chemical constituents or phytochemicals (Ghazanfar, 1994). Phytochemicals are responsible for metabolic activities and used for defense purpose. They are produced by specific biochemical pathways, which occur inside the plant cells. Due to their physical characteristic and chemical properties these chemical constituents are highly beneficial to the mankind and most importantly in primary health care. The first chemical substance isolated from plants was benzoic acid (Figure 1) discovered in 1560. The potent pain-killer, morphine (Figure 1) was separated from the dried latex of Papaver somniferum L. (opium) by F. W Serturner (1783-1814). The bioactive molecule, ephedrine (Figure 1) is one of the most potent plant based medicine and was isolated from Ephedra sinica (Ma huang). It was used in the form of various salts to combat bronchial asthma (Tang & Eisenbrand, 1992). Reserpine (Figure 1) an indole alkaloid, obtained from the roots of Indian plant, Rauwolfia serpentina, was used to treat hypertension. Taxol (Figure 1) is a diterpenoid first isolated from the stem bark of pacific Yew, Taxus brevifolia and used against ovarian for the treatment of ovarian cancer.

1. Introduction

Mounting pressure of teeming population and shrinking land resources have necessitated the production of increasingly more food per unit area. Insect pests, diseases and weeds are important biotic constraints inflicting 20-25% loss in agriculture production. Synthetic pesticides have of course, played a very significant role in restricting many pest problems. However, their indiscriminate has been its toll by creating several problems like pesticide resistance in insects, contamination of food with toxic residues, resurgence of pests and deleterious effects on non target organisms. The number of effective pesticides available is dwindling day by day further having less choice among which to choose. These ill effects have arised interest in alternate methods of plant protection.

The extent to which various plant species are utilized for pest control depends, not only on their effectiveness against target pests, but also on socio-economic, environmental and policy considerations. Keep these in mind the present study on the efficacy of botanicals against Spodoptera litura was carried out.

Ethnopharmacology and Traditional medines use natural resources as medicines. They have developed the knowledge of this through the generations of traditions and experiences. Therefore the protection of this knowledge and protection of natural resources both remain critical. Bioprospecting involves sustainable, ethical and legal processes to access natural resources and use them judicially so that the business needs do not pose any threat to nature and to people.

One important consideration is assuring the preservation of traditional knowledge, particularly oral knowledge, within communities, for the potential benefit of others. Developing mechanisms to document traditional knowledge, such as national inventories, can be beneficial in identifying cases where conservation or cultivation is needed, based on an understanding of existing natural resources used for medicinal purposes. Decisions about access and ownership of information can also be made (including those relating to intellectual property rights, where appropriate) once methods have been found to collect and preserve traditional knowledge. On the other hand, the livelihood of traditional healers may depend on maintaining the secrecy of their healing methods - one argument for consulting with traditional healers as part of the process of planning and executing the collection, storage and testing of traditional medicines.

Extending some form of intellectual property protection to traditional knowledge, of which traditional medical knowledge is a significant part, is being actively debated in WIPO and other international forums. Existing intellectual property regimes can offer protection for purified compounds (produced by scientists and companies) isolated from a natural medicine. But traditional medicines, whose method of action may not be well understood, and which often consist of mixtures of different active substances, may not meet criteria for patentability. The principal impetus for international debate is a desire to compensate the holders of traditional knowledge appropriately for the use of their knowledge by others, or to prevent so-called “biopiracy” where knowledge or genetic materials are used without the consent of the holder. China and Kenya are examples of countries that have modified their laws in an effort to accommodate traditional medicine. It is not intended that the Commission should enter into this debate, except in so far as aspects of it may be relevant to its terms of reference, in particular stimulating innovation in traditional medicine with the purpose of expanding access to affordable medicines.

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.

The main purpose of herbal drug technology is to convert botanical materials into medicines with a focus on standardization and quality control with proper integration of modern scientific techniques and traditional knowledge. The use of chromatographic techniques and marker compounds to standardize botanical preparations has limitations because of their variable sources and chemical complexity. DNA-based molecular markers have utility in fields like taxonomy, physiology, embryology, genetics etc. DNA based techniques have widely been used for authentication of plant species of medicinal importance. Pharmacognosy mainly addresses the quality related issues using routine botanical and organoleptic parameters of crude drugs, and chemoprofiling assisted characterization with chromatographic and spectroscopic techniques. The new pharmacognosy includes all the aspects of drug development and discovery where the biotechnology driven applications play an important role. Current focus on chemotype-driven fingerprinting and related techniques require integration with genotype-driven molecular techniques so that an optimal characterization of botanical materials is possible. This review provides a brief account of various DNA based technologies that are useful in genotyping and quick identification of botanicals with suitable examples.

Introduction
Animal sector is growing very fast particularly in India as people’s purchasing power is increasing day by day. With increase in income, people’s dietary habit is shifting towards more animal origin products. The use of antibiotics for animal food production is growing very fast and India is the 4th largest country in the world regarding consumption of antibiotics in veterinary and animal husbandry sector. There is urgent need of AMR mitigation strategies to reduce the effect on human and animal health. As per the World Bank AMR can affect the global GDP upto 3.8% by 2050. Most of the countries in the world are very cautious regarding antibiotic residues and rejecting the livestock products which lead to economic loss. Many world agencies/forum such as CodexAMR Guidelines, UN Political Declaration on AMR Guidelines, Global Leaders Group on AMR reiterated judicious use of antibiotics. All-India Network Project on AMR inLivestock and Fisherieswas started in India to reduce the effect of AMR by educational awareness, adoption at field level, surveillance system and diagnosis but it didn’t meet out the desired outcome. Capacity building programmes in the field of AMR by training, reform in curriculum, collaboration from different fields of professionals can mitigate the harmful effect of AMR. According to the Food and Agriculture Organization (FAO), education and training play pivotal roles in translating knowledge into practical implementation, warranting the inclusion of Antimicrobial Use and AMR as integral components in professional education, postgraduate training, and continuing education within the food and agricultural sectors Training Strategies It involve Antimicrobial Stewardship training for healthcare professionals, public awareness campaigns, farmer education, and veterinary training on responsible antimicrobial use. Other critical elements include fostering collaboration through professional networks, building surveillance capacity, utilizing AI-driven tools for better decision-making, and promoting hygiene and infection prevention to reduce infection incidence and the need for antibiotics.
Training for Healthcare Professionals
There is substantial evidence that both healthcare workers and members of the public have knowledge gaps about appropriate use of antibiotics and mechanisms of antibiotic resistance. Surveillance is a prerequisite to estimate the magnitude of the AMR burden and to establish any intervention strategies, such as antimicrobial stewardship. In order to launch successful intervention approaches through cooperative efforts from different stakeholders (e.g., international agencies, human and veterinary medicine sectors, agriculture and animal production industries, and consumers), the existing knowledge gap needs to be addressed first.

Cardiac ultrasound is also known as echocardiography and it has a very important place in the diagnosis of cardiac diseases of the humans as well as animals. Echocardiography is an imaging of the heart using ultrasound. The diagnosis of congenital heart diseases, shunts, complex malformations, valvular heart diseases, different cardiomyopathies, endocarditis, pericardial effusion, cor pulmonale and cardiac tumours has been possible by cardiac ultra ultrasound that was otherwise difficult.

Phytochemicals are the individual chemicals from which the plants are made. Phytochemical is simply a word that means plant chemicals. Plants have the ability to synthesize mixtures of structurally diverse bioactive compounds with multiple and mutually potential therapeutic effects. The plants have the capacity of manufacturing the secondary products. Phytochemicals with antioxidant properties tend to be brightly colored because they contain chromophores, ie, a series of alternating single-bonded and double-bonded carbons. Isoprene is often the building block of such units. The darkest green vegetables contain the most chlorophyll, and vegetables with the most chlorophyll require the most antioxidants.

1. Introduction
Animal experimentation remains a cornerstone of scientific research in many fields, from medical advances to pharmaceutical testing. However, the ethical treatment of laboratory animals has become a point of significant concern worldwide. India, through the Committee for the Control and Supervision of Experiments on Animals (CCSEA), ensures that ethical standards are maintained in research institutions. Established under the Ministry of Environment, Forests, and Climate Change (MoEF&CC), CCSEA is tasked with overseeing the use of animals in experiments, ensuring that procedures adhere to ethical guidelines and international norms. CCSEA’s guidelines are in line with international standards set forth by organizations such as the International Council for Laboratory Animal Science (ICLAS) and the European Union (EU), but they are tailored to address local needs and conditions. The guidelines, which focus on the welfare of laboratory animals, emphasize the ethical review process, animal care, and the importance of minimizing animal suffering. They also stress the need for researchers to implement the 3Rs—Replacement, Reduction, and Refinement—to minimize the use of animals and ensure humane practices in research. This chapter explores the CCSEA guidelines in detail and compares them with animal welfare regulations from around the world, including the United States, European Union, Japan, and Australia. By understanding the similarities and differences between these regulations, researchers and policymakers can ensure the ethical treatment of laboratory animals on a global scale.
2. Objectives of CCSEA Guidelines
The main objective of the CCSEA guidelines is to ensure that laboratory animals are treated ethically and humanely while also supporting the validity and scientific rigor of research. The guidelines encourage researchers to prioritize animal welfare while minimizing unnecessary suffering. Specifically, CCSEA focuses on:
1. Ethical Treatment of Animals: Ensuring that animals are housed in suitable conditions, given adequate veterinary care, and provided with a comfortable environment to reduce stress.
2. Justification of Animal Use: Ensuring that animals are used only when necessary, and that their use is scientifically justified.
3. Pain and Stress Minimization: Ensuring that experimental procedures cause the least pain, distress and suffering possible.

This chapter describes the cells, tissues and organs responsible for immune defence. It examines lymphocytes, macrophages, dendritic cells, neutrophils, mast cells, natural killer cells and plasma cells, emphasizing their respective functions. Primary and secondary lymphoid organs—including bone marrow, thymus, bursa of Fabricius, spleen, lymph nodes and mucosa-associated lymphoid tissue—are discussed. The chapter also covers lymphocyte maturation, antigen presentation, phagocytosis, immune-cell surface markers and the organization of lymph-node regions. Positive and negative selection of T lymphocytes and the formation of memory cells are included. Together, these topics explain how immune components develop, communicate and coordinate effective responses against infectious agents.