eChapters

1. Introduction
All glassware used for bacteriological work must be meticulously cleaned and sterilized. Used and soiled glassware should be thoroughly washed to remove dirt and solid residues before drying and undergoing sterilization.
2. Cleaning of New Glassware
Borosilicate glass (e.g., Pyrex) and factory-washed soda glass require only standard washing before use. However, new, unwashed soda glass needs additional treatment to neutralize alkali content.
• Place the items (e.g., test tubes, Petri dishes, and flasks) in a suitable container, cover with a 1% tri-sodium phosphate solution, and bring to a boil.
• Remove the items, rinse with tap water, and immerse them in a 1% hydrochloric acid solution to neutralize alkalinity.
• Rinse thoroughly with tap water, followed by distilled water. Allow the glassware to dry before sterilization.
3. Cleaning of Used Glassware and Other Apparatus
• Autoclave all glassware contaminated by microorganisms (121°C for 15–20 minutes). This liquefies solid media, facilitating removal.
• Rinse the empty glassware with tap water, then warm water (50–55°C).
• Soak in a detergent solution (e.g., 1% washing soda or tri-sodium phosphate solution) and scrub with a brush.
• Rinse thoroughly with tap water to remove detergent traces, followed by distilled water.

Introduction
Ocular micrometer and stage micrometer are used to measure the size of microorganisms. The Ocular micrometer consists of a circular disk of glass which has graduations engraved on surface. A stage micrometer or objective micrometer has scribed lines on it that are exactly 0.01mm (10 micrometer) apart. Ocular micrometer is calibrated for the objective first. To calibrate the ocular micrometer for a given objective, the two scales are superimposed and determined how many of the ocular gradation coincide with one graduation on the stage micrometer scales. Thus, the distance between the graduations on the ocular micrometer can be determined and so the size of the organism can be determined.
Materials
1. Ocular micrometer
2. Stage micrometer
3. Microscope
4. Stained slides of yeast or bacteria

Introduction
This method consists of mixing the microorganisms in a small amount of India ink or nigrosine and spreading the mixture over a clean slide. These two pigments are not really bacterial stain and do not penetrate the microorganisms. These pigments make the background dark thus making organisms transparent against dark background. This method is useful for determining cell morphology and accurate size as no shrinkage of cells due to heat.
Materials
1. Bacterial culture (B. subtilis)
2. Microscopic slides (Clean, Grease Free)
3. Inoculating needle
4. Cedar wood oil
5. Microscope
6. Xylene

5.1 Purpose

To employ a standard protocol to perform agarose gel electrophoresis.

5.2 Principle

Agarose gel electrophoresis is the most effective way of separating DNA fragments of varying sizes ranging from 100 bp to 25 kb. Migration rate of DNA molecule depends on various factors like 1) size of DNA molecule; 2) agarose concentration; 3) DNA conformation 4) voltage applied, 5) presence of ethidium bromide, 6) type of agarose and 7) electrophoresis buffer. After separation, the DNA molecules can be visualized under UV light.

Disease    Prescribed test    Alternative test

This chapter introduces hybridoma technology and its application in producing monoclonal antibodies. It explains the fusion of antibody-producing B lymphocytes with immortal myeloma cells, selection in HAT medium and the roles of HGPRT and polyethylene glycol. The survival and cloning of hybrid cells and the advantages of monoclonal antibodies are considered. The chapter also discusses tissue and organ transplantation, including autografts, isografts, allografts and xenografts. The immunological mechanisms of acute graft rejection, donor histocompatibility antigens and the involvement of T lymphocytes are addressed. These questions connect laboratory antibody production with transplantation immunology and its diagnostic, therapeutic and research applications.

Veterinarians will be often required to mention the specific animal husbandry terms for various purposes like writing certificates for various schemes, postmortem, attending court of law in veterolegal cases etc. Hence this chapter aims in refreshing the vets with the common terms associated with different domestic animals and animal husbandry activities. 

The basic knowledge of normal physiological values in different animal species is essential for selection of the animal, diagnosis of any abnormality if any and its correction. The chapter provides the normal physiological values viz temperature, pulse, respiration etc in different domestic animals.

The different biochemical values serve as an indicator of the correct health status, viz important organ functions,  the alteration in values if any, helps in identifying the specific organ damage and exact diagnosis of the  abnormal health condition or disease. Some of the important biochemical values of the animals, which may assist in diagnosing the specific disease  conditions are listed in this chapter.

The  information on normal reproductive traits in domestic animals like calving interval, age at first calving etc, is essential for getting the maximal reproductive efficiency of the animal by the farmer. It also helps in improving the reproductive efficiency of the animals by different required means viz nutrient/mineral supplementation, antibiotic, hormonal therapy etc.

Among various immunodiagnostic tools enzyme immunoassay becomes increasingly popular for the detection of humoral immune response (seroconversion) or microbial antigens for the purpose of diagnosis. Enzyme Linked Immunosorbent Assay (ELISA) is widely used in studies of infectious diseases.Despite several merits of ELISA, it has some limitations such as requirement of ELISA reader to obtain accurate measurements. To overcome this, the dot immunobinding assay was developed .In Dot Immunoassay nitrocellulose membrane is used as an adsorbent matrix instead of plastic surface.The antigen is dotted onto the nitrocellulose membrane and the membrane then incubated with test antibody, and secondly with enzyme conjugated antibody directed against the first antibody.In case of positve
reaction, (specific antigen –antibody reaction) color dot is developed against the white background at the site of antigen deposition.In case of negative reaction, no color development takes place at the site of antigen deposition .
Principle
• In direct ELISA antibody is directly attached to solid phase and targeted by added antigen (detecting antigen).
• These added antigens are targeted by antispecies antibodies linked to enzyme termed as conjugates.
• The bound enzyme is developed by the addition of substrate /chromogen, then stopped and finally read using a spectrophotometer

The table gives the approximate date of parturition, based on the date of breeding/mating in the year as a easy ready reference table.