Pathology for Toxicologists: Principles and Practices of Laboratory Animal Pathology for Study Personnel

305 Pages·2017·8.75 MB·English

by  Elizabeth McInnes

Table of Contents
Title Page
copyright
List of Contributors
Preface
Chapter 1: An Introduction to Pathology Techniques
1.1 Animal Considerations
1.2 Necropsy
1.3 Lung Inflation with Fixative
1.4 Fixation
1.5 Making Glass Slides
1.6 Special Histochemical Stains
1.7 Decalcification
1.8 Immunohistochemistry
1.9 Tissue Crossreactivity Studies
1.10 Electron Microscopy
1.11 In Situ Hybridisation
1.12 Laser Capture Microscopy
1.13 Confocal Microscopy
1.14 Image Analysis
1.15 Digital Imaging
1.16 Spermatocyte Analysis
1.17 Good Laboratory Practice
1.18 Inhalation Studies
1.19 Continuous-Infusion Studies
1.20 Carcinogenicity
1.21 Biologicals
1.22 The Pathology Report
1.23 Conclusion
References
Chapter 2: Recording Pathology Data
2.1 What is a Pathology Finding?
2.2 Standardisation of Pathology Findings
2.3 ‘Inconsistencies’ in Pathology Recording
2.4 Blind Review
2.5 Historical Control Data: Pros and Cons
2.6 The Use of Peer Review in Pathology
References
Chapter 3: General Pathology and the Terminology of Basic Pathology
3.1 Cellular Responses to Insults
3.2 Inflammation
3.3 Circulatory Disturbances
3.4 Disorders of Tissue Growth
3.5 Tissue Repair and Healing
3.6 Neoplasia
3.7 Immune System
References
Chapter 4: Common Spontaneous and Background Lesions in Laboratory
Animals
4.1 Rats
4.2 Mice
4.3 Dogs
4.4 Minipigs
4.5 Non-Human Primates
4.6 Rabbits
4.7 Experimental Procedures
4.8 Causes of Death in Rats and Mice
4.9 Conclusion
References
Chapter 5: Target Organ Pathology
5.1 Skin
5.2 Eye
5.3 Gastrointestinal Tract
5.4 Liver
5.5 Respiratory System
5.6 Urinary System
5.7 Lymphoreticular System
5.8 Musculoskeletal System
5.9 Cardiovascular System
5.10 Endocrine System
5.11 Reproductive System
5.12 Central and Peripheral Nervous System
5.13 Ear
References
Chapter 6: Clinical Pathology
6.1 Clinical Pathology in Study Phases and Good Laboratory Practice
6.2 What is Measured in Clinical Pathology?
6.3 Haematology
6.4 Coagulation
6.5 Clinical Chemistry
6.6 Urinalysis
6.7 Acute-Phase Proteins
6.8 The Biomarker Concept
6.9 Reference Intervals
6.10 Instrumentation, Validation and Quality Control
6.11 Data Analysis and Interpretation
6.12 Reporting
6.13 Food Consumption and Body Weight (Gain)
6.14 Organ Weights
6.15 Examples of Typical Clinical Pathology Profile Changes in
Toxicologic Clinical Pathology
6.16 Microsampling
6.17 Conclusion
Acknowledgments
References
Chapter 7: Adversity: A Pathologist's Perspective
7.1 LOAEL, NOEL and NOAEL: Definition
7.2 Adversity
7.3 Determining Adversity using Pathology Findings: Factors to
Consider
7.4 Communicating NOAEL in Toxicity Studies
7.5 Conclusion
References
Chapter 8: Limitations of Pathology and Animal Models
8.1 Limitations of In Vivo Animal Models
8.2 Efficacy/Disease Models as Toxicology Models
8.3 Limitations of Efficacy/Disease Models as Toxicology Models
8.4 Limitations of Pathology within In Vivo Toxicology Models
8.5 Managing Risk Associated with Subjectivity and the Potential for
Pathologist Error
References
Glossary
Index
End User License Agreement
List of Illustrations
Chapter 1: An Introduction to Pathology Techniques
Figure 1.1 Necropsies or post mortem examinations on experimental
animals such as this mouse are a fundamental part of toxicological
pathology.
Figure 1.2 Post mortem imbibition of bile pigment in the mesenteric fat
tissue adjacent to the gall bladder. Photograph taken from a bovine
necropsy.
Figure 1.3 Artefacts induced at necropsy include inclusions of foreign
material into the issue, such as plant material (*) during brain removal.
Figure 1.4 Production of glass slides suitable for histopathological
analysis.
Figure 1.5 Tissue trimming and placement in a cassette.
Figure 1.6 The blocking sheet.
Figure 1.7 Small pieces of tissue (*) from a previous cassette
superimposed on another tissue.
Figure 1.8 Machine for tissue dehydration, clearing and impregnation
with paraffin wax.
Figure 1.9 Rotary microtome.
Figure 1.10 H&E staining.
Figure 1.11 Quality control.
Figure 1.12 Phosphotungstic acid haematoxylin (PTAH).
Figure 1.13 Growth-hormone IHC stains positive cells in pituitary.
Figure 1.14 Phospholipidosis is characterised by lamellar bodies (*),
which are visible in this electron micrograph of a rat heart.
Chapter 2: Recording Pathology Data
Figure 2.1 How a pathology term is created.
Figure 2.2 Chronic progressive nephropathy (CPN) is a common and
well-recognised background finding in rats. The lesion progresses from
occasional basophilic tubules in the early stages to multiple different
morphological findings in the later (chronic) stages. A pathologist who
lumps this lesion will record different grades of CPN throughout the time
course of the lesion whereas a splitter will describe the different findings
seen at each stage.
Chapter 3: General Pathology and the Terminology of Basic Pathology
Figure 3.1 Severe cell injury is not difficult to recognise. Here, the pus
exuding from this ruminant brain clearly indicates that the tissue is not
normal. This is called ‘purulent meningitis’.
Figure 3.2 Yellow fatty change in an enlarged ruminant liver.
Figure 3.3 Infarcts (*) in a dog kidney, showing a sharp demarcation
between normal (red-brown) tissue and necrotic (yellow) tissue.
Figure 3.4 Coagulative necrosis (*) in a ruminant kidney.
Figure 3.5 Liquefactive necrosis in a ruminant brain.
Figure 3.6 Caseous necrosis in the centres of ruminant liver abscesses.
Figure 3.7 Fat necrosis in ruminant mesenteric fat tissue.
Figure 3.8 Dry gangrene in the black ear tips of a pig.
Figure 3.9 Fibrinoid necrosis in beagle pain syndrome, showing bright
pink material in the blood vessel wall.
Figure 3.10 Apoptosis (*) in a mouse lymph node, showing cell
shrinkage, chromatin condensation, cytoplasmic blebs and phagocytosis
of apoptotic bodies by adjacent macrophages.
Figure 3.11 Clinical signs of inflammation.
Figure 3.12 Major actions of the exudative phase of acute inflammation.
Figure 3.13 Cells involved in inflammation: neutrophils, macrophages,
eosinophils, mast cells and basophils.
Figure 3.14 The complement system.
Figure 3.15 Types of inflammation.
Figure 3.16 Fibrinous inflammation around a ruminant heart.
Figure 3.17 Formation of fibrous tissue (scar tissue) (*) in the scars of
old infarcts in a dog kidney.
Figure 3.18 Simultaneous tissue destruction and repair in inflammation.
Figure 3.19 Multinucleate giant cell.
Figure 3.20 Functions of inflammation.
Figure 3.21 Hyperaemia on the surface of a ruminant brain.
Figure 3.22 Petechiae on the surface of a canine heart.
Figure 3.23 Hydrothorax is the accumulation of fluid in the thoracic
cavity.
Figure 3.24 Conversion of fibrinogen to fibrin during clotting
(haemostasis).
Figure 3.25 Thrombosis in a canine large blood vessel.
Figure 3.26 Epistaxis in a dog.
Figure 3.27 Cell growth responses.
Figure 3.28 Thickened whitish scar at the edge of an ulcerative lesion.
Figure 3.29 Interrelationship between different systems in inflammation.
Chapter 4: Common Spontaneous and Background Lesions in Laboratory
Animals
Figure 4.1 Squamous cyst (*) in a mouse spinal cord.
Figure 4.2 Bluish thymus within the thyroid follicles of a mouse.
Figure 4.3 Haematopoiesis (*) in the liver.
Figure 4.4 Thickening of the glomerular basement membranes and
periglomerular fibrosis (*) in an ageing mouse, indicative of CPN.
Figure 4.5 Thickened mucosa filled with dilated gastric glands,
indicating adenomatous hyperplasia of the glandular stomach.
Figure 4.6 Enlarged nuclei (*) in the cells of the liver.
Figure 4.7 Enlarged mouse spleen (*) at necropsy.
Figure 4.8 Loss of hair and ulcerative skin lesions in a C57Bl/6 mouse.
Figure 4.9 Lymphoma, manifesting in enlarged submandibular lymph
nodes and enlarged thymus.
Figure 4.10 Pituitary tumour in a female Sprague Dawley rat.
Chapter 5: Target Organ Pathology
Figure 5.1 Fur/hair loss or thinning (alopecia) in a mouse.
Figure 5.2 Acanthosis (epidermal thickening) in a young rat pup.
Figure 5.3 Necrosis, erosion and ulceration of the skin in the preputial
gland area of a male mouse.
Figure 5.4 Petechiae on the skin of a nude mice.
Figure 5.5 Squamous cell carcinoma in the dorsal skin above the scapula
in a rat.
Figure 5.6 Conjunctivitis in a mouse.
Figure 5.7 Whitish opacification of the eye due to the presence of a
cataract.
Figure 5.8 Squamous papilloma in the mouth of a rat.
Figure 5.9 Squamous cell carcinoma in the stomach of a rodent.
Figure 5.10 Chronic colitis (*) in a mouse.
Figure 5.11 Polyp in the colon of a rat.
Figure 5.12 Leiomyosarcoma extending from the ileum outer surface in a
rat.
Figure 5.13 Yellow areas (*) on the reddish-brown surface of the liver,
indicating liver necrosis and inflammation.
Figure 5.14 Jaundice/icterus in a mouse, visible as a generalised yellow
colour of the abdominal muscles.
Figure 5.15 Hepatocellular adenoma (*) in the liver of a mouse.
Figure 5.16 Distended, air-filled murine stomach and intestines, due to
obstruction of the airflow due to inflammation of the nasal turbinates.
Figure 5.17 Slightly raised white foci on the lung surface, indicating the
presence of macrophage aggregates.
Figure 5.18 Bronchiolaveolar adenoma (*) in the lung of a mouse.
Figure 5.19 Shrunken, wrinkled, pale white rat kidneys at necropsy,
indicating fibrosis and chronic interstitial nephritis.
Figure 5.20 Yellow left kidney with raised abscesses (*), indicating
pyelonephritis in a mouse.
Figure 5.21 Urinary stones in the bladder of a mouse.
Figure 5.22 Hydronephrosis in both kidneys of a mouse.
Figure 5.23 Haematuria (*) in a mouse urinary bladder.
Figure 5.24 Severe enlargement of the spleen (*) due to lymphoma in a
mouse.
Figure 5.25 Severely enlarged submandibular, axillary and inguinal
lymph nodes in lymphoma in a mouse.
Figure 5.26 Enlarged parathyroid gland (*) in renal failure in a dog.
Figure 5.27 Small red raised areas in the lung of a dog, indicating the
spread of a haemangiosarcoma from the spleen.
Figure 5.28 Pituitary adenoma (large, dark red mass at the base of the
cranial cavity) (*) in a rat.
Figure 5.29 Enlargement of the thyroid (*) in a rat.
Figure 5.30 Phaemochromocytoma in an adrenal adjacent to the kidney
in a rat.
Figure 5.31 Testicular atrophy (*) in the left testis of a mouse.
Figure 5.32 Ovarian cyst (*) in a mouse.
Figure 5.33 Mammary gland fibroadenoma in a rat.
Chapter 6: Clinical Pathology
Figure 6.1 The three phases in clinical pathology.
Figure 6.2 The main materials for toxicologic clinical pathology testing
are anticoagulated whole blood for haematology (e.g. in EDTA), serum
collected after centrifugation of clotted whole blood for clinical
chemistry and Wright–Giemsa-stained blood smears for microscopic
evaluation and validation of quantitative and qualitative aspects of red
and white blood cells and platelets. The arrow points to the feathered
edge, where platelet clumps or atypical cells can preferentially be seen.
Figure 6.3 Microhaematocrit tubes with haemolysis, lipaemia (small fat
layer after centrifugation; arrow) and marked icterus in the plasma layer.
Such discolourations can be accidental or treatment-related and can
interfere with analytical measurements.
Figure 6.4 Increased numbers of reticulocytes indicate active
haematopoiesis in the bone marrow, a physiologic response to blood loss
due to moderate or excessive blood sampling, haemorrhage or
haemolysis. (a) Traditionally, reticulocytes are visualised by vital stains,
such as new methylene blue, which make the reticular pattern of remnant
ribosomes and RNA visible. (b) In the haematology analyser,
reticulocytes are identified by their remnant RNA (in the absence of a
nucleus) (x-axis), with most immature reticulocytes on the far-right side.