eChapters

A decoration, interpretation, or visual explanation of a text, concept, or process that is intended to be included in published media including posters, flyers, magazines, books, teaching materials, animations, video games, and films is called an illustration.

Using Illustrations

We can comprehend information better when it is shown. Illustrations were one of our first forms of communication, long before any kind of contemporary writing. In reality, new discoveries of cave drawings from more than 35,000 years ago show that humanity have been using images to communicate for a very long time. This did not occur by accident; according to study, over 90% of the information we process is visual, and a significant portion of our brain is dedicated to visual processing.

Benefits of using Illustrations

The sensors present over remote sensing satellites are designed to collect energy or radiance from the varied surface features having wide range of radiance i.e. from features like water bodies having very low radiance to features like sand or snow having very high radiance. But, most of the time, the study area may not have all these features; therefore, radiance values may remain confined to a narrow range. This reduces the contrast of the image, thereby decreases its visual quality. In this context, image contrast enhancement techniques are used for effective visualization and display of remote sensing image on computer screens or hard copy so that maximum amount of information can be derived through its visual interpretation.

Contrast enhancement of a digital image is a technique to modify the original grey levels of input image to the entire available range of the grey levels. This increases the contrast between various features in the image which improves the visual quality of original image. Contrast enhancement can be done by both linear and non-linear methods. Some of the commonly used contrast enhancement techniques with their implementation in R software are discussed in this chapter.

We need following packages for performing this exercise in R. raster package is required for importing of raster image and for performing contrast enhancement. RStoolbox package is also required for contrast enhancement as well as for visualization of raster data with ggplot2 package. rasterVis package is mainly used in this exercise for plotting histogram of the raster data.

The information of image is perfectly accepted by eyes when the image will be in convenient form. Always pictures accurately detail about information related to their position, sizes and inter-related with each other is objects. Actually images portray positional information which can be recognized as objects. Human eyes are exact example for differentiating that information from images to innate visual and mental ability.

Abstract

Before 1960’s, UAE was a poor developing country and almost there were no real signs of civilization. During the 1970’s two important things happened: H. H. Sheikh Zayed Bin Sultan Al Nahyan, the president of UAE, and the other Emirates rulers united together and established the United Arab Emirates and considerable oil findings were discovered. The UAE Leadership has proved to have deep future vision; in three decades the country jumped from a relatively backward scattered emirates to a well developed and civilized country.

Development is very obvious : from tents and sandy tracks to tower buildings and highways, from desert to green belts of palm trees and vegetables in all parts of the country, and most important, education, healthcare and all other services became the best in the region. Urban development was so sharp; someone who left the place 10 years ago can hardly know his location and services in all sectors became the best at the international level. The oil money was properly invested in development.

UAE Air Force Space Reconnaissance, used satellite and aerial images for the past 20 years and successfully utilized images from various sources including aerial photographs, Russian satellites images, SPOT and Landsat

I. Dairy Farm Equipments

19.1 Introduction

Nano technology or Nano Science is a new branch of research though nano materials or Nano Structures are not new. In nature nano particles or nano materials are abundant .The idea of manipulation, production, controlling size and physico - chemical properties and application as well as making devices out of these nano materials is the subject of Nano Science & Nano Technology. As the name suggests, the domain of Nano starts from 10-9 m but especially the effect of nano structures exists mostly in the 1-100 nm regime where quantum - mechanical behiviour is prevalent. Molecules with more than 10 atoms fall into the nanometer range  and scientists were not able to visualize individual molecules and nano scale objects until the invention of some very powerful microscopes. Working in Nano domain requires two basic facilities -

Almost all imaging techniques employed in human medicine are being followed in canine medicine but radiography and ultrasonography ( abdominal and cardiac) are the most commonly used clinical diagnostic techniques. Computed tomography, magnetic resonance imaging, and nuclear techniques are also being used in the teaching establishments.

Radiography

Use of radiography (Figure 42) as a diagnostic technique is wide spread in canine and feline medicine across the world. It is considered the first step in evaluating the skeleton, thorax and abdomen. For evaluating the abdomen, radiography is considered as complimentary to sonography. Canine practice is more like pediatrics requiring assistant to control the patients (dog sand cats). No anesthesia or sedation is generally required for radiography. But if the patient has painful lesion such as fracture or luxation, suitable anesthesia / sedation should be used for making the animal cooperative during radiography for better visualization of the lesions.

IMF : Short for International Monetary Fund, referee and, when the need arises, rescuer of the world’s financial system. The IMF was set up in 1944 at bretton woods, along with the world bank, to supervise the newly established fixed exchange rate system. After this fell apart in 1971–73, the IMF became more involved with its member countries’ economic policies, doling out advice on fiscal policy and monetary policy as well as microeconomic changes such as privatisation, of which it became a forceful advocate. In the 1980s, it played a leading part in sorting out the problems of developing countries’ mounting debt. More recently, it has several times co-ordinated and helped to finance assistance to countries with a currency crisis.

Immunity, active : Resistance developed in response to stimulus by an antigen (infecting agent or vaccine) and usually characterized by the presence of antibody produced by the host.

Immunity, herd : The resistance of a group to invasion and spread of an infectious agent, based on the resistance to infection of a high proportion of individual members of the group. The resistance is a product of the number susceptible and the probability that those who are susceptible will come into contact with an infected person.

Immunity, passive : Immunity conferred by an antibody produced in another host and acquired naturally by an infant from its mother or artificially by administration of an antibody-containing preparation (antiserum or immune globulin).

Defination

Dislocation of carpometarcarpus and tarso metatauses along with upper fracture of MC and MT bone (Fig. 1 and Fig.2). It is considered as difficult fracture and may not be healed appropriately by the usual procedure of lower level fracture repair method. Due to constant movement of fracture ends its treatment should be similar to middle level fracture repair method.

Materials Required

Thick padded cloth

Adhesive gum (fevicol)

Netted bamboo splint

Thomas splint

Purpose

As the immobilisation of upper third part of radius ulna is very difficult ,the purpose of the immobilisation tech. Is to fix the upper third of radius-ulna with the posterior half of ring of Thomas splint with bandages are very important for achieving no movement of fracture fragment during healing.

Indication

Correction of upper third fracture of radius-ulna by Thomas splint application indicated in higher level fracture.

Objectives

Immobilization of fracture of mandible.

Requirements

‘U’ Plate 4 Pins Electric drill Orthopaedic hammer Pin cutter

Procedure

Under local infiltration of an anesthetic & parenteral sedatives the 4 pins are drilled 2 on either side of the fracture site from one side mandible to other side mandible. During drilling the pin, the holes, importance should be given for the passage of the pin through prefix of the drill hole in metal frame

The fracture end should come accurate apposition during pinning procedure & the symmetry of the fractured mandible should be as norma! as possible.

Sterilised antiseptic gauge bandages are covered, the pin between frame and the body. The pins are retained in position either with nut-bolt or by passing a plastic tube after which the hole of the plastic tube is filled with resin.

To know the method of immobilization of the limb after repair of the transected gastrocnemius tendon (tendo achiles).

Procedure

The immobilization procedure of limb after gastrocnemius tendon repair is done in 2 stages.

1^ST Stage

Immobilization of limb for avoiding movement at tendon suture site.

It is done by placing the sutured area within a pre- fabricated metal frame so as to keep the limb undisturbed at the suture site.

The frame is given in Figure 1.

The purpose of addition of the metal frame is to immobilize the limb keeping the fetlock and foot away from in a flexed manner to avoid weight bearing during healing period.

The limb is immobilized with netted bamboo splint (Fig. 2).

The immobilized limb is placed within the metal frame keeping the fetlock and foot in a flexed manner

Aim: To know the method of immobilization of the limb after repair of the flexor tendon.

Procedure

The immobilization procedure of limb after flexor tendon repair is done in 2 stages.

Definition: Mandible forms the skeleton of lower jaw and comprises of a body and two rami. The two halves of this bone fuse incompletely at the mandibular symphysis, situated at the midline within the body.

Indication: Immobilization of fractured mandible, minimally or non displaced fractures with an intact opposite hemimandible, vertical ramus fracture.

Anasthesia: Local anaesthesia for large animals. A20 gauze 2.5 cm needle is used to deposit 5-1 Omi of lidocaine at the exit site and into the canal.

Infection refers to the invasion of harmful microorganisms into the body and causing disease. The most important function of the immune system is to protect the body of the host against various viruses, bacteria, fungi and parasites. The grounds of immune surveillance stand on the basis of innate and specific immunity that build up against the evasive mechanisms of the disease causing organisms. Infectious disease is result of a complex interaction between  the host and the microbes. Evasions of protective immunity by the parasites critically determine the survival and pathogenicity of the microbes. The invasion of the body by harmful microorganisms is called infection. Infections are dangerous in any part of the body. Microorganisms spread vary easily into the blood stream and cause blood poisoning which is called septicemia.

Infectious diseases are the result of a complex interaction between the host and the microbes. Evasions of protective immunity by the parasites critically determine the survival and pathogenicity of the microbes. The invasion of the body by harmful microorganisms is called infection.

A. Answer the following.

Q.1.     How lysozyme kills Gram + and Gram – bacteria ?
Ans.     Lysozyme kills Gram + bacteria by splitting cell wall proteoglycans and Gram – bacteria in conjunction with complement.

Q.2.     Describe the different forms of leprosy ?
Ans.     Leprosy occurs in two different forms (1) tuberculoid (2) lepromatous. Tuberculoid leprosy occurs in 80% cases. Tuberculoid leprosy patients mount a strong CMI response. These patients have few organisms in their tissues and therefore few lesions. The lesions are infiltrated with macrophages and patients show a positive delayed type hypersensitivity reaction to I/D

The study of the immune system, in its various aspects, is called immunology. Immunology is the branch of bioscience which deals with the defense mechanisms including all physical, chemical and biological properties of the organism that help it to combat its susceptibility to foreign agents and microorganisms. Immunology includes the development and functions of various components of immune system by which the body responds, destroys or neutralizes foreign agents or pathogens. It includes the study at cellular as well as molecular level. Immunology is an interdisciplinary subject which incorporates and requires the knowledge of many branches of biology like microbiology, biochemistry, molecular biology and clinical medicine.

Immunity

Immunity is the resistance of body against extraneous etiological factors of disease, which is afforded by the interaction of chemical, humoral and cellular reactions in body. This is an integral part of the body without, which one cannot think of life. During the process of evolution, nature has provided this defense mechanism in the bodies of all living creatures particularly of higher animals and man, that protects them from physical, chemical and biological threats. It can be classified as natural or paraspecific and acquired or specific immunity.

Natural/paraspecific immunity

There are some species which are resistant to particular diseases due to presence of natural resistance against them e.g. horse, pig, cat are resistant to canine distemper virus; dogs are resistant to feline panleucopenia virus, chickens are resistant to anthrax. Even within species, there is natural resistance that protects some individuals while others are susceptible e.g. Indian deshi cattle Zebu (Bos indicus) is quite resistant to piroplasmosis in comparison to Bos taurus. Besides, there are the mechanisms or barriers in body provided by nature.

Introduction

Anatomy of the immune system in birds differs from that in mammals. Immune cells are present in primary lymphoid organs (PLO) and secondary lymphoid organs (SLO). In birds, thymus and bursa of Fabricius (cloacal bursa) are primary lymphoid organs. Secondary lymphoid organs include the spleen, bone marrow, Harderian gland, aggregates of lymphoid tissue in various organs; i.e. in the gut called gut-associated lymphoid tissue (GALT); in bronchi called bronchial- associated lymphoid tissue (BALT). SLO contains aggregates of lymphocytes and antigen presenting cells. Birds do not have lymph nodes.

In PLO, the lymphoid cells are processed for differentiation and maturation. T lymphocytes differentiate and mature in the thymus, whereas B lymphocytes mature in bursa of Fabricius. Functional lymphocytes move from PLO to SLO. Like mammals, birds defend the body against pathogens through innate immunity (non-adaptive immunity) and adaptive or acquired immunity. Under innate immunity system, physical barriers present in birds play their important role preventing the entry of pathogens into the body. Physical barriers are feathers, skin, aggregates of lymphoid cells in various organs and ciliary defence in the respiratory system. Phagocytosis is first defence for pathogens entering the body, evading the physical barriers. Phagocytic cells include mainly heterophils and macrophages. Natural killer cells (NK cells), thrombocytes and complement also help in innate immunity.

Adaptive immunity is a particular defence which develops when the innate immunity is inadequate to impart protection. Adaptive immunity is antigen specific. T cell, B cell and macrophages play a major role in adaptive immunity. Adaptive immunity is brought about by antibodies (humoral immunity) or also by cells (cell-mediated).

In animals with a long gestation period, such as domestic animals, the acquired immune system is fully developed at birth, but cannot function at full adult levels for several weeks. The complete development of the acquired immunity depends on antigenic stimulation. The proper development of B cells and B cell antigen receptor (BCR) diversity requires clonal selection and antigen driven cell multiplication. Thus, the newborn animals are vulnerable to infection for the first few weeks of life. Hence, the temporary assistance in defending themselves at this time is provided by the mother in the form of antibodies and possibly T cells.
 
Development of the Immune System

The thymus is first lymphoid organ to develop, followed closely by the secondary lymphoid organs. B cells appear soon after the development of spleen and  lymph nodes, but antibodies are not usually found until late in foetal life, if at all. The ability of fetus to respond to antigens develops very rapidly after the lymphoid organs appear, but all antigens are not equally capable of stimulating fetal lymphoid tissue. The immune system in a series of steps, each step permitting the foetus to respond to more antigens. These steps are driven by a gradual increase in the use of gene conversion or somatic mutation to antibody diversity. The ability to mount cell mediated immune (CMI) responses develops at the time as antibody production. Data from humans suggest that T cell receptor diversity is limited in the fetus and neonate and that cytokine production may be low. This may simply be due to their lack of to foreign antigens.

Depressed immune responses have been observed frequently in cancer patients. Cancer and other stress are known to affect different components of the immune system. Cancer treatment is relying mainly on surgery, anti-cancer chemotherapy and radiation therapy. Most chemotherapeutic agents are responsible to myeloid toxicity, which leads to leukemia and immunosuppression affecting both humoral and cell-mediated immunity. This is a very serious disadvantage of chemotherapy besides they are cytotoxic drugs.

Diagnosis of parasitic infections is routinely done with conventional parasitological techniques as described earlier. However, when the parasitaemia is too low to be detected by the conventional methods as in subclinical infections and/or carrier animals advanced diagnostic tools are used for the early, sensitive as well as specific diagnosis.

Serological techniques

Diagnostic techniques used mostly in laboratories need various reagents and equipments to perform the test. However, a diagnostic kit is the extrapolation of the same method by packing it in a user friendly format, accompanied by all or most of the reagents required to perform the test and can also be done outside the laboratory or under field conditions.

Diagnosis of parasitic infections is routinely done with conventional parasitological techniques as described earlier. However, when the parasitaemia is too low to be detected by the conventional methods as in subclinical infections and/or carrier animals advanced diagnostic tools are used for the early, sensitive as well as specific diagnosis. Serological techniques Diagnostic techniques used mostly in laboratories needs various reagents and equipments to perform the test. However, a diagnostic kit is the extrapolation of the same method by packing it in a user friendly format, accompanied by all or most of the reagents required to perform the test and can also be done outside the lab or under field conditions. Various immunodiagnostic kits are available which are based on different diagnostic methods like Enzyme-linked immuno sorbent assay (ELISA). Complement fixation test (CFT), Agar gel immunodiffusion test (AGID), Immunofluorescent tests (IFT), Latex agglutination tests (LAT) etc. ELISA is the most commonly used test system and numerous kits based on ELISA are available commercially.

Immunoblotting or Western blotting is a technique employed to observe proteins separated through gel electrophoresis. After the proteins migrate from the gel to a nitrocellulose or PVDF membrane due to an electrical current, the membrane is exposed to antibodies tailored for the desired target. Subsequently, secondary antibodies and detection reagents are utilized to visualize the proteins of interest on the membrane.

1. Preparation of tissue homogenate

The initial phase of sample preparation involves extracting proteins from their origin. Typically, proteins are extracted from cells or tissues through a process called lysis. This procedure disrupts the cell membrane to separate proteins from the insoluble components of the cell. Different lysis buffers are available for sample preparation in Western blotting and they differ in the potency of their detergents used to liberate soluble proteins.

Introduction

Immunology deals with the body defence system consisting of various cells and biomolecules that prevents entry of foreign infectious and toxic agents by different mechanisms. The mammalian immune system operates in two ways like innate or non specific immunity and adaptive or acquired or specific immunity. Innate part constitutes the physical (skin, hair, tears and mucous secretions) containing enzymes that prevents primarily entry of pathogen. It is the first line of defence action, very fast but not adequate. Adaptive form constitutes defensive cells and secreted immunoglobulins. It takes time to develop effectively, especially when the body sees the organism for the first time. Immunoglobulins (Ig) or antibodies are a group of proteins produced from B lymphocytes and plasma cells in response to the presence of any foreign substances. They are 5 basic types like M, A, G, E and D each with specific function. The system can produce antibody molecules, directed to about 1 million different kinds of antigens. All immunoglobulins have a similar basic structure.

The well-being of a person depends on his immune system. Any deficiency in any component of immune system may lead to defects in the normal functioning of his body and makes him more susceptible to diseases. The immunodeficiency may be either in specific components (B cells or T cells) or nonspecific component (phagocytes and complement) of the immune system.

The wellbeing of a person depends on his immune system. Any deficiency in any component of the immune system is called immunodeficiency and it may impair the normal functioning of his body and makes the person more susceptible to infections.

A. Answer the following.

Q.1.    Which protein is coded by gag gene in HIV ?
Ans.     It codes for protein that form the core (p24 and p17).

Q.2.     Which protein is coded by pol gene in HIV ?
Ans.    It codes for a protein that cleaved into 4 enzymes. 

Q.3.     Which protein is coded by env gene in HIV ?
Ans.    It codes for protein that is cleaved into two proteins (gp120 and gp41).

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.

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:

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.

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.