VETERINARY CLINICAL PATHOLOGY AND LABORATORY DIAGNOSIS
authored by: R.S.Chauhan & Desh Deepak Singh
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Clinical Pathology and laboratory Diagnosis are often required to be in one location to assist veterinarians and disease investigation experts in their routine work. The development of rapid and specific diagnostic tools for various animal diseases is an important area of focus for clinicians. Traditional methods for diagnosing infectious diseases involve isolating, growing, and identifying the infectious agent from clinical samples, which requires well-equipped and well-staffed pathological laboratories. These methods can take days to weeks and may not provide an early diagnosis, which can be crucial for managing the disease. While isolation and identification of the microorganism can provide an unambiguous diagnosis, the success rate is limited. An alternative approach is to detect seroconversions to particular microorganisms, which can be diagnostically important. Enzyme immunoassay has become a popular tool for detecting humoral immune responses or microbial antigens for diagnostic purposes. Additionally, this book provides regulatory and safety requirements for the first time for veterinary disease investigation. Field veterinarians can use this information in their daily routine work to diagnose diseases, identify causative agents, and recommend effective therapeutic and preventive measures to control livestock and poultry diseases.
Prof RS Chauhan – COWPATHY MAN
MVSc, PhD, FNAVS, FSIIP, FIAVP, Diplomat ICVP, PDCR, ACPPM, OCTT, MBA
EX- Director ICAR-IVRI, JD CADRAD, Director IBT, National Fellow, Advisor WHO. Member, Animal Welfare Board of India; Member, CPCSEA ( Govt. of India); Chairman, Scientific Advisory Committee, ICMR-NARFBR.
Born on September 10th, 1958 in a farmer's family, Dr. Chauhan completed his Bachelors (80.3%) with honours, Master (85.8%) and Doctoral degree (90.0%) in Veterinary Sciences with specialization in veterinary pathology from G.B. Pant University of Agric. & Tech., Pantnagar. He served the country in various capacities including Assistant Professor/ADIO (1983-96), Associate Professor (1996-1999), National Fellow (1999-2004) and Joint Director (CADRAD), IVRI, Izatnagar(2004-2009), Director & Vice-Chancellor(2009), ICAR-IVRI and Campus Director, IBT, Patwadangar (2009-2013). During his tenure as academician and scientist, he has written 112 books including 35 manuals and 1 monograph very popular among the students world over. He contributed 99 chapters in different books and published 235 research and 59 review papers. Besides, he participated in International / National Conferences and presented 178 papers. He is life member of 15 scientific bodies and has been in several executive committee such as Chairman, Panch Gavya Professionals’ Club; President, Cow Therapy Society; Secretary-General, Society for Immunology and Immunopathology; Vice President, Indian Society of Veterinary Educators; Zonal Secretary, Indian Association of Veterinary Pathologists and Joint Secretary, Indian Virology Society, President, Dr. J.L. Vegad Foundation, Expert Member Research Group, Committee for Certification of Pathologists, Vice-President, IAVP; Registrar, ICVP, etc.
Based on his contributions and scientific achievements, he has been awarded with several prizes, medals and honours including Best Young Scientist Award (1992), IAAVR Award (1996), National Fellow Award (1999), Fellow NAVS (2000), Fellow SIIP (2001), K.S. Nair Memorial Award (1999), Vigyan Bharti Award (2000), Dr. C.M. Singh Trust Award (2002), Dr. Rajendra Prasad Award (2002), Shri Ramlal Agrawal National Award (2000), Best Paper Award SIIP (2003), Best Paper Award IAVA (2001, 2002, 2003), Best Teacher Award (2004) by GBPUAT, Fellow, IAVP (2006), Gopal Gaurav (2007), Bharat Excellence Award (2007), Diplomat, ICVP (2008), Intas-ISVE Best Veterinary Scientist Award (2008), Best Academician Award (2012), Moropant Pingle Go Sewa National Award (2015), Outstanding Scientist Award (2019), Research Excellence Award (2020), Indo Asian-Claude Bourgelat Distinguished Innovative Scientist Award-2020 in Animal Immunopathology and MAN OF COWPATHY at GADVASU Ludhiana etc. in recognition of his research and teaching endeavor. He has been inducted in many national and international scientific/advisory committees and boards including Member, WHO/IPCS Committee on Environmental Health Criteria. Dr. Chauhan added many new including Enterotoxaemia, Pyometra, ETEC infection in camels, isolated camel pox virus, developed rapid diagnostic test DIA for the first time in India. He reported role of cell mediated immunity in rotavirus infection in calves. His pioneer work includes immunopathology due to pesticides, heavy metals, mycotoxins and nanoparticles. Dr. Chauhan developed a new method using MTT dye for detection of CMI response. He has scientifically validated Panchgavya and named it as “Cowpathy”.
He have been recognized internationally as visiting Professor, University of Wageningen and Advisor, WHO. Dr. Chauhan guided more than 50 scholars for their Masters and Doctoral research; most of them are placed as Professors, Scientists, Officers in Indian Army, banks and industry in India and abroad. At present he is working as Head Veterinary Pathology, Chairman Scientific Advisory Committee ICMR-NARFBR, Member AWBI and CPCSEA (Govt of India).
Desh Deepak Singh, M.V.Sc., Ph.D., Professor & Head,Department of Veterinary Pathology, Uttar Pradesh Pandit Deen Dayal Upadhyay Pashu Chikitsha Vigyan, Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura-281001, Uttar Pradesh, India
"1. Examination of Blood
2. Examination of Blood and Serum for Biochemical Attributes
3. Examination of Bone Marrow
4. Examination of Urine
5. Examination of Feces
6. Examination of Milk
7. Examination of Skin Scrapings
8. Examination of Synovial Fluid
9. Examination of Cerebrospinal Fluid
10. Examination of Semen
11. Clinical Microbiology
12. Cytopathology
13. Necropsy
14. Histopathological Techniques
15. Immunological and Serological Techniques
16. Immunopathological Techniques
17. Toxins and Poisons
18. Molecular Biological Techniques
19. Investigation of an Outbreak
20. Diagnosis of TB, JD and Sexually Transmissible Diseases (STDs)
21. Validation of A Diagnostic Test
22. Management of Veterinary Clinical Pathology Laboratory: Safety, Accident And First AID
23. Biosafety in Veterinary Clinical Pathology Laboratories
24. Quality Management System (Iso 9001: 2008) in Veterinary Clinical Pathology Laboratory
25. Competence Accreditation (Iso 17025) of Veterinary Clinical Pathology Laboratory
26. Good Laboratory Practice in Veterinary Clinical Pathology Laboratory
27. Clinical signs, Pathological Lesions and Samples Required for Laboratory Examinations
28. Miscellaneous Information for Diagnosis and Prevention of Diseases
The need of a suitable literature for students of Veterinary Sciences on disease investigation was felt since long. Many times it has been observed that Clinical Pathology and Laboratory Diagnosis requirements for disease investigation are required to be at one place so that the Veterinarians and disease investigation experts find it useful for their routine disease investigation work. The need of rapid and specific diagnostic tool(s) for detecting a wide variety of viral, bacterial, fungal, toxic, metabolic and parasitic diseases of animals has always been an area of considerable importance for clinicians.
• General considerations • Collection of blood • Storage • Examination of blood • Coagulation time • Bleeding time • Determination of movement of parasites under wet film • Total erythrocyte count • Total leucocyte count • Differential leucocyte count • Hemoglobin • Erythrocyte sedimentation rate • Hematocrit value • Mean corpuscular volume • Mean corpuscular hemoglobin • Mean corpuscular hemoglobin concentration • Erythrocyte fragility test
• General considerations • Blood glucose • Total serum protein • Serum albumin • Serum globulin • Serum gamma globulin • Serum calcium • Serum inorganic phosphorus • Serum magnesium • Serum cholesterol • Serum creatinine • Serum urea • Serum chloride • Serum sodium • Serum potassium • Serum Glutamic Oxaloacetic Transaminase • Serum Glutamic Pyruvic Transaminase • Serum alkaline phosphatase
• General considerations • Indications • Limitations • Techniques of bone marrow examination • Cell types • Cellular composition of bone marrow • Morphology of the cells • Interpretation of bone marrow examination
• General considerations • Collection • Preservation • Examination • Physical examination • Chemical examination • Microscopic examination • Organized sediment • Unorganized sediment • Cultural examination
• General considerations • Collection of feces • Storage • Examination of feces • Gross examination • Microscopic examination • Examination of feces for flukes • Examination of feces for blood • Culture of feces for nematode larvae • Sporulation of coccidia.
• General considerations • Collection of milk sample • Storage • Examination of milk • Physical examination • Chemical examination • Cytological examination • Bacteriological examination
• General considerations • Examination of skin scrapings for mange • Examination of skin scrapings for fungi • Cultural examination General Considerations The skin scrapings are examined for the diagnosis of skin diseases, including parasitic, fungal and bacterial infections and infestations.
• General considerations • Collection of synovial fluid • Examination of synoival fluid • Physical examination • Chemical examination • Cytological examination • Microbiological examination • Examination for immune complexes • Miscellaneous examination • Inflammatory conditions of joints • Non-infectious arthritis • Infectious arthritis • Immune complex mediated arthritis
• General considerations • Indications • Collection of cerebrospinal fluid • Sub-occipital puncture • Lumbar puncture • Examination • Physical examination • Chemical examination • Cytological examination • Microbiological examination
• General considerations • Volume • Colour • Reaction • Concentration of spermatozoa • Motility • Live and dead count • Microbiological examination
• General considerations • Direct examination • Culture methods • Some important tests used in identification of bacteria • Characteristics of some organisms • Gram positive cocci • Gram positive rods • Gram negative rods • Mycobacteria • Leptospira/spirochete • Rickettsia • Chlamydia • Mycoplasma • Fungi • Virus • Antibacterial sensitivity testing
• General considerations • Scope • Exfoliative cytopathology • Interventional cytopathology (Biopsy) • FNAC • Imprint cytopathology • Crush smear cytopathology
• General Consideration • Post-mortem examination of large animals • Post-mortem examination of poultry • Steps in post-mortem examination • Writing of post-mortem report • Collection, preservation and dispatch of specimens for laboratory diagnosis
• General considerations • Scope • Microscopic examination techniques • Smear • Vital staining • Routine histopathological techniques of sectioning • Collection of tissues • Fixation • Washing • Dehydration • Clearing • Impregnation • Casting of blocks • Trimming • Section cutting
• General considerations • Agglutination • Slide agglutination test • Tube agglutination test • Capillary tube agglutination test • Plate agglutination test • Hemagglutination test • Hemagglutination-inhibition test • Coagglutination test
• 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
• General considerations • Cyanide poisoning • Nitrate poisoning • Nitrite poisoning • Oxalate poisoning • Strychnine poisoning • Arsenic poisoning • Lead poisoning • Thalium poisoning • Ergot poisoning • Carbon monoxide poisoning • Phenothiazine poisoning • Pesticide poisoning
• General considerations • Polyacrylamide gel electrophoresis (PAGE) • SDS-polyacrylamide gel electrophoresis (SDS PAGE) • Nucleic acid hybridization • Radio labelled probes • Biotin labelled probes • DNA finger printing • Polymerase chain reaction (PCR) • Reverse transcriptase polymerase chain reaction (RT PCR)
• General considerations • First hand information and constitution of a team • Gathering the disease information from records and the owner • Animal data • Previous illness • Present disease • Proforma for animal examination report • Observation of team • Logical clinical examination of animals • Disease investigation kit
• General considerations • Tuberculosis • Johne’s disease • Brucellosis • Infectious bovine rhinotracheitis • Campylobacteriosis • Tichomonosis
• General considerations • Steps in validation • Step1- Feasibility studies • Step 2- Assay development and standardization • Step 3- Determining assay performance characteristics • Step 4- Monitoring validity of assay performance • Step 5- Maintenance and enhancement of validation criteria • Measurement of uncertainity of a diagnostic test
• General considerations • Ethics for veterinary diagnostic laboratory personnel • Safety measures in the laboratory • Accidents • First aid • Acid burns • Alkali burns • Poisoning • Burns caused by heat • Injuries caused by broken glass • Contamination by infectious materials • Bodily damage by electric shock • Precautionary measures
• General considerations • Biosafety levels-1 and -2 • Biosafety level-3 • Biosafety level-4 General Considerations Biosafety is a combination of laboratory practices, techniques, equipments and other facilities appropriate for the protection of hazards posed by infectious agents handled in the laboratory. There are guidelines for maintaining the biosafety in laboratory at various levels described as under:
• General considerations • Quality management principles • Objectives of 9001:2008 1. Scope 2. Normative reference 3. Terms and definitions • Main clauses and sections of ISO 9001:2008 4. Quality management system 5. Management responsibility 6. Resource management 7. Product realization 8. Measurement, analysis and Improvement
• General considerations • Purpose and goals of laboratory • Standards, guides and references • Accreditation for competence • Identification of scope • Test methods • Benefits of ISO17025 accreditation • Processing of application
• General considerations • Purpose • Scope • Five pillars of GLP • Good laboratory practice principles • Test facility organization and personnel • Quality assurance programme • Facilities • Apparatus, materials and reagents • Test systems • Test and reference items • Standard operating procedures • Performance of the study
• Nutritional diseases • Viral diseases • Prion, Mycoplasmal, Rickettsial, Chlamydial, Spirochaetal and Bacterial diseases • Fungal diseases • Parasitic diseases • Toxic conditions • Miscellaneous conditions
• Epidemiological definitions • List of tests for international trade • OIE listed diseases • Solutions and reagents • Age and body weight of animals • Rectal temperature of animals • Normal values of blood vascular system • Vaccination schedule
A Abdominal 126, 130, 131, 132, 141, 149, 334, 488, 496, 504, 507, 508, 509 Abdominal cavity 130, 131, 132 Abomasum 132, 144, 149, 269, 473, 477, 487, 504, 507 Abortion/ metritis 155, 511 Abscess 46, 58, 93, 95, 98, 99, 100, 143, 144, 146, 151, 488, 490, 492, 498, 409 ABTS 202, 292, 293, 294, Acetone 52, 54, 108, 177, 237, 241, 242, 271, 272, 279, 282, 290, 292, 297, 298, 299, 300, 300, 301, 302, 303, 377, 386 Acetonitrile 282, 283, 300, 304 Actinobacillus 116, 491 Actinomycosis 151, 492 Acute inflammation 13, 95
clinical pathology, laboratory diagnosis, veterinarians, disease investigation, diagnostic tools, animal diseases, infectious diseases, pathological laboratories, seroconversions, enzyme immunoassay, humoral immune responses, microbial antigens, regulatory requirements, safety requirements, veterinary disease investigation, therapeutic measures, preventive measures, livestock diseases, poultry diseases