eChapters

Definition

IDD (immunodeficiency diseases) occur when there is a failure of the immune system to protect the host from infectious organisms or the development of cancer.

IDD are disorders of immune system that result in immunological deficiency. Immunodeficiency could be from defects in

i. B and T- lymphocytes

ii. Granulocytes

iii. Complement

This chapter focuses on the structure, classes, properties and biological functions of immunoglobulins. It introduces the basic antibody molecule, including heavy and light chains, variable and constant regions, antigen-binding sites and Fc fragments. The characteristics of IgG, IgM, IgA, IgE and IgD are examined in relation to their distribution and immune functions. Questions cover primary and secondary immune responses, placental transfer, mucosal protection, allergic reactions, complement activation and agglutination. The chapter also addresses antibody affinity, avidity, isotypes, allotypes, idiotypes, class switching and Bence-Jones proteins. It provides essential knowledge of how antibodies recognize antigens and contribute to immune protection and diagnosis.

• General considerations

 • Agglutination

 • Slide agglutination test

 • Tube agglutination test

 • Capillary tube agglutination test

 • Plate agglutination test

 • Hemagglutination test

 • Hemagglutination-inhibition test

 • Coagglutination test

Immunological Causes of Male Infertility

Immune mediated male infertility is an auto-immune disease.Spermatozoa are first produced at puberty and isolated from the systemic circulation by a structural barrier maintained by Sertoli cell tight junctions within the testis and the uninterrupted epithelial cell lining of the excurrent ductal system. Sperm production occurs well after the development of immunological tolerance to self antigens; therefore sperm specific components are viewed as foreign by the immune system. Whether spermatogonia, the primordial cell in spermatozoa differentiation, is also antigenically foreign is not established. What is known , is that immunization with sperm elicits a highly reproducible immune response that can impair fertility.

Antisperm antibodies are found in about 10-15% of infertile men. Immunity only to those antigens present on the surface of viable, motile sperm will impair fertility. Antibodies to internal sperm antigens, due to an immune response to sperm degraded in the genital tract or to cross reacting microbial antibodies present in the sera of many fertile individuals , are of questionable relevance to infertility. A number of spermatozoa antigens have been identified .

The evolution process took millions of years to develop human and animals on the top of the hierarchy. It has moved forward due to ability to survive the various deadly diseases as for the survival of any leaving creature, it have to overcome the threat posed by the pathogens surviving in its environment and most of the time in its close vicinity. These microbes are smarter than us and are the best example of the survival of the fittest. To overcome these, nature has provided defense system to all leaving creatures that neutralize the invasion of pathogens or undesired microbes. The concept is based on the detection of self and nonself molecules.

The pathogenicity of an organism varies form mild attack to severe fatal attacks. The host has many mechanisms to resist or to avoid the pathogens apart from immune systems. One or few of these will be activated when the pathogen enters into the body. Following are some of the listed natural host defense mechanisms.

I. IMMUNOLOGY OF MALE REPRODUCTIVE SYSTEM

There are two basic functions of the immune surveillance of man and animals with respect to sperm immunity during spermatogenesis and fertilization process: support of the elimination of the vast majority of 'waste "spermatozoa that fail to fertilize and preservation of the tiny minority of surviving spermatozoa that are able to fertilize the oocytes. The primordial forms of spermatozoa arising in the seminiferous tubules of testes are preserved by the blood-testes barrier which is formed by sertoli cells ringing the semineferous tubules (Hunter, 1989). During maturation in the epidydimis, the spermatozoa acquire their antigenic properties. Simultaneously, the spermatozoa are coated with some proteins of seminal plasma having the immunosuppressive activity toward lymphocytes and macrophages involved in the immune reactions. The seminal plasma proteins accompany the migration of spermatozoa to the female genital tract (Matousek, 1985).

Introduction

To improve the growth and health status, addition of antibiotics in the poultry feed was a common practice in last decade. Currently, in developed countries it is banned due to fatal consequences like development of antimicrobial resistant organism pool in the gut, persistence of antibiotic residue in the food chain that can reduce the food safety level. Therefore, various alternatives are now a day thought to use as poultry feed additives. Immunomodulaors are presently receiving wide attention as feed additives.

The term ‘immunomodulation’ is generally used to describe the manipulation of immune system which consists either increase (immunostimulation) or decrease (immunosuppression) in the magnitude of the immune system function. The chemicals or biological substances producing this phenomenon are known as ‘immunomodulators’. Specific immunomodulation involves the change in the immune response with a particular antigen (e.g. vaccination). Whereas, non-specific immunomodulation alters the immune response in such a way so that it become sensitive to a wide range of antigens. Generally this kind of non-specific immunomodulators are used as poultry feed additive to increase the resistance against the wide range of infections. The principal components of the immune system targeted for immunomodulation include T cells, B cells, monocytes, macrophages, granulocytes and natural killer cells (Dalloul and Lillehoj, 2005).

The concept of immunonutrition has been developed in humans and animals mainly in relation to its impact on reinforcement of the immune system and its ability to resist disease. However, in the case of immunology of reproduction, different (even opposite) mechanisms  of immune system modulation are likely to be involved since the main objective is a local inhibition or the attenuation of the immune system toward the protection of the embryos. Most of the studies investigating the role of feeding and /or specific nutrients have used nutrient deficiency models to identify the mechanisms involved. During recent years, an increasing number of substrates have been recognized for their immunomodulating function. Macro and micronutrients have been identified as so-called immunonutrients. Although informative, such an approach might not apply to a concept of immunonutrition for reproduction when the dietary levels of a given nutrient will have to cover both the metabolic and immunomodulation needs for optimal reproduction.

1. Introduction
Aquaculture has rapidly emerged as a crucial industry to meet the growing global demand for seafood, contributing significantly to food security and economic development. However, as aquaculture operations expand and intensify, fish health management has become increasingly challenging. The close confinement of fish in high-density farming systems creates ideal conditions for the spread of infectious diseases. Stressors such as overcrowding, fluctuating water quality, and improper nutrition further exacerbate the susceptibility of fish to pathogens. As a result, disease outbreaks in aquaculture are more frequent and can lead to severe economic losses due to reduced productivity, increased mortality, and the cost of treatments. In the past, antibiotics and other pharmaceutical treatments were widely used to manage fish diseases. While these methods have been effective in controlling infections, they come with several significant drawbacks:
• Antibiotic resistance: Overuse of antibiotics has contributed to the rise of antibiotic-resistant bacteria, a major public health concern that can affect not only aquatic organisms but also humans through the food chain.
• Residues in fish products: Antibiotics and other chemicals may leave harmful residues in fish, posing potential health risks to consumers and limiting the export potential of farmed fish products to markets with stringent regulations.
• Environmental contamination: The use of chemicals in aquaculture can lead to the contamination of surrounding water bodies, affecting wild fish populations and the broader ecosystem.
Due to these limitations, there is a growing demand

• Allergic Tests

 • Single intradermal test

 • Tuberculin test

 • Johnin test

 • Double intradermal test

 • Comparative test

• Mallein test

 • Immunofluorescence Techniques

 • Direct method

• Indirect method

 • Immunoperoxidase Techniques

 • Detection of antigen

Materials and Reagents Required
1. Fixative- cold acetone or methanol
2. Ethanol – 70%, 90% and 100% for rehydration and dehydration
3. Xylene for clearing
4. Washing buffer (phosphate buffer saline)- PBS
5. Hydrogen peroxide – for endogenous peroxidase blocking
6. Normal horse serum or any other serum: 2.5% horse serum or any other serum can be used to block non-specific epitopes.
7. Ki67 mouse primary antibody
8. Secondary antiboy kit - Impress Excel kit (Cat no. MP-7601; Vector Laboratories, NY, USA) or a similar kit from other vendors like Biogenex (Figure 22; left panel).
9. Peroxide substrate kit—Immpact NovaRed (cat no. SK-4850; Vector Laboratories, NY, USA) for developing a red color signal (Figure 22; right panel).

Background
Immunostaining is a technique used to visualize specific proteins or antigens within cells. In milk somatic cells, immunostaining is employed to identify and quantify various cell types, study their functional characteristics, and diagnose diseases. Immunostaining can be used to identify and characterize different cell types based on the expression of specific surface markers. For example, CD45 is a leukocyte marker, while cytokeratins are markers for epithelial cells. In addition to the research on mammary gland biology, immunostaining is a valuable tool for studying the biology of the mammary gland. It can be used to investigate the expression of genes involved in milk production, cell differentiation, disease processes and even pregnancy detection. For example, galectin-3-binding protein (LGALS3BP) is a gene with elevated expression in the PBMCs of early pregnant buffaloes (Sharma et al., 2023). In this chapter, we have investigated the protein expression of LGALS3BP in milk somatic cells to determine if immunopositive cells are present in milk. Apart from this, stem cells in milk can be identified using markers like CD133, CD73, CD44,and other cell surface markers (Coni et al., 2023).

Background
Immunocytochemistry is the immunostaining of a specific cell or its components in a cell smear. It is a technique to visualize a specific protein or an antigen in cells based on an antigen- antibody interaction (Figure 10). The site of antibody-antigen binding can be visualized by color development (enzymechromogen system) or fluorescence. The macrophages can be visualized in the smear of peripheral blood mononuclear cells by immunocytochemistry using antibodies against macrophages. We need to use a light or fluorescent microscope to detect color or fluorescence, respectively.

In the study of immunology, several techniques are used. These techniques are called immunotechniques. These techniques are used not only in immunology but are the methods for research and diagnostic purpose in many fields such as microbiology, biochemistry, endocrinology, hematology and in pathological laboratories.

Many techniques are used in immunology to detect specific proteins (antigens) or antibodies in biological samples. These techniques are called immunotechniques. These methods are used for research and diagnostic purpose in many fields such as microbiology, biochemistry, endocrinology, hematology and in pathological laboratories. The techniques can be used to estimate proteins, drugs, hormones and toxic substances in a sample. Immunological techniques are highly specific, precise and reliable. These methods are simple and safe. Antigen and antibody reaction is highly specific in nature and this forms the base of the immunological methods. There are many immunotechniques which are used for the qualitative and quantitative estimation of antigens and antibodies in a sample. Some of commonly used techniques in immunology are:

Indicators for Impact Assessment-meaning and Concept

Indicators for impact assessment: Indicators are common tools to determine the performance and then the result of the technology/extension/communication interventions. They measure the accomplishment of the project’s goals and targets. Criteria of indicators should be defined during the formulation stage, but they often need to be specified in greater detail during implementation. The indicators should measure intangible and non-tangible changes, particularly in projects that value factors such as personal and social development.

Indicators for impact assessment
Meaning and concept of Indicator
The term indicator is often used interchangeably with data, targets, standards for evaluation, and even various modes of data collection, which undoubtedly inhibits a productive dialog on the subject. Such confusion is unwarranted, since the term has a precise connotation- in the literature on monitoring and evaluation of development. Simply stated, indicators are variables designed to measure change in a given phenomenon or process. They are analytical tools which facilitate the measurement of change and provide summary data for project design, implementation, and evaluation. Indicators are defined as
“specific (explicit) and objectively verifiable measures of changes or results produced by an activity or intervention” (United Nations: 1985; p. 37). A few observations about indicators can be useful in further clarifying their nature. First, as the above definition implies, indicators provide quantitative data which can be analyzed using statistical techniques to produce estimates of a phenomenon. In cases when only qualitative information is gathered, it should be converted into numerical data for the purposes of analysis and presentation. Second, an indicator can capture one or more dimensions of a phenomenon or process depending upon the purpose of the study. We can, for example, construct an indicator for urbanization based on a wide range of variables such as number of school, higher educational institution, road connectivity, number of financial institution, health infrastructure, income etc. On the other hand, one can use each of the above items as a distinct indicator. Indicators focusing on different dimensions of a subject are difficult and more costly to construct. Third, several indicators can be used to capture the same phenomenon.