Pathology Flashcards Weeks 13-18
In terms of analyzers, why are medical labs not good for veterinary use?
Medical labs are unlikely to have the correct animal add-on software for their analyzers and differentials may be wrong due to differences in cell size and morphology between animal and human blood
In terms of technicians and pathologists, why are medical laboratories not useful for veterinary use?
Technicians in medical labs are unlikely to be trained to deal with animal samples, and human pathologists will not discuss cases with you
In terms of tests and reference ranges, why might medical laboratories not be appropriate for veterinary use?
They will not have reliable reference ranges for haematological and biochemical parameters in animals, and tests are likely to be inappropriate for animal use
What is vital to fill in when submitting samples to a lab?
Signalment, Clinical Signs and History
How should specimens be packaged when submitting samples to a lab?
The specimens should be packaged in such a way that they will not break or leak and are ideally surrounded by soft packing material such as bubble wrap
What are 4 reasons for why it is advantageous to use Veterinary Practice laboratories?
Excellent service for vet clients, Rapid Turnaround, Reliable Results, They have Vet Pathologists
Who is responsible for interpretation of laboratory results?
The practitioner is responsible, even when interpretive comments are provided by a laboratory pathologist.
What is required when interpreting laboratory results?
A table of references is required in order to identify any abnormalities
What do reference ranges represent?
The distribution of test results in a healthy population. The range is considered to be the mean (+/- 2 standard deviations)
Why is repeat testing required?
If a biochemical test of say 15 tests is performed, there will be a greater than 50% chance of finding one or more values outside the reference range. Therefore, variations from normal are not always straightforward.
What is the quality assurance programme?
A standardized set of protocols from handling samples to reporting results which helps ensure the reliability, precision and accuracy of test results.
What are quality control programmes?
A set of processes and instruction for a machine or diagnostic test that ensure the results produced are as reliable and accurate as possible
What is internal quality control?
Where samples of known value are run and the results are comapred to the known values to make sure they are within acceptable limits
What is external quality control?
Where results of tests on unknown controls are compared with “consensus” means calculated from the results of all participating laboratories
What are the five parameters that indicate how good a test is?
Accuracy, Precision, Sensitivity, Specificity and Predictive Values
What is the accuracy of a test?
Assesses how close the test result is to the actual test
What is the precision of the test?
Assesses how repeatable the test result is when testing the same sample
What is the Sensitivity of the test?
The measurement of the frequency with which a test result will be positive in an animal that actually has the disease
What is specificity?
The measurement of the frequency with which a test result will be negative in an animal that does not have the disease
What is a predictive value of a test?
Based on the sensitivity and specificity of a test but also factors in the vets pre-judgement of the likelihood of the disease being present.
What are the 3 major cell types present in the blood, and how much of the body mass is water?
Erythrocytes, Leukocytes and Platelets - 60-70% of the total body mass is water
Describe the purpose of the haematopoietic system
The haematopoietic system manufactures the cellular components of blood and is widely distributed throughout the body
Describe the role of bone marrow with regards to the haematopoietic system
Main source of erythrocytes, granulocytes, monocytes and thrombocytes. Also stores iron for heme synthesis, and is the source of stem cells for T and B-lymphocytes.
Describe the role of the liver with regards to the haematopoietic system
Major haematopoietic organ during the first half of embryonic life. Function is gradually overtaken by bone marrow, but still has potential for haematopoiesis in adults.
Describe the role of Lymph Nodes with regards to the haematopoietic system
Produces B and T-lymphocytes and plasma cells. Participates in antibody production and cell mediated immunity.
Describe the role of the spleen with regards to the haematopoietic system
Produces T and B-lymphocytes and plasma cells, stores erythrocytes and iron, removes aged and abnormal erythrocytes - also degrades haemoglobin.
Describe the role of the thymus with regards to the haematopoietic system
Controls differentiation of bone marrow derived stem cells into T-lymphocytes
Describe the role of Macrophages of the Monocyte Phagocytic System with regards to the haematopoietic system
Macrophage line sinusoids in spleen, liver, lymph nodes, bone marrow and destroy aged or abnormal erythrocytes and haemoglobin. Also stores iron.
Where does erythropoiesis occur and what is involved?
In the bone marrow and involves the differentiation of a pluripotential stem cell into an erythroid cell, and after several forms it becomes a mature erythrocyte
In response to eryrthropoietin (EPO) what happens to erythroid stem cells?
Erythroid stem cells differentiate into a rubriblast and begin to produce haemoglobin and oter proteins.
Over a period of 3-4 days nucleated erythroid precursor cells progress into what?
A series of divisions and maturation processes that result in the production of 8-16 rubicytes from each rubriblast
What is the final stage in erythroid maturation, and when do they become ‘reticulocytes’?
Metarubicytes are the final nucleated stage in maturation and once the nucleus is extruded, they become reticulocytes
When is nucleus extrusion triggered?
When the erythrocyte reaches a critical level of haemoglobin content
How long does it take for reticulocytes to be released from bone marrow?
~7 days from the time of initial stimulation of stem cells by EPO
What are the 3 major pools of erythrocyte storage?
Erythrocyte precursors (mainly in bone marrow), Blood (Mainly Mature Erythrocytes and Spleen (Mainly Mature Erythrocytes)
How does the kidneys regulate erythropoiesis?
The kidneys produce erythropoietin, which stimulates stem cells to differentiate into rubriblasts and promotes haemoglobin synthesis
What minerals are involved in Erythropoiesis?
Vitamin B12, Vitamin B6 and Folic Acid, Iron, Copper and Cobalt are essential for erythrocyte production
What hormones are involved in Erythropoiesis and what are their effects?
Androgens, which stimulate erythropoiesis and Oestrogens, which have an inhibitory effect
What is extravascular haemolysis?
The process by which old or damaged erythrocytes are removed from circulation
What happens to old red blood cells during extravascular haemolysis?
They become less pliable and are phagocytosed by macrophages lining sinusoids in the spleen, liver and bone marrow
During extravascular haemolysis, what is disassembled and what is released?
Haemoglobin is disassembled and iron is released. The iron is either stored intracellularly as haemosiderin or transported via transferring back to the bone marrow for re use in erythropoiesis. Billirubin also released.
What is erythrocytosis?
An increase in PCV, erythrocyte count or haemoglobin concentration above the normal range
What is relative erythrocytosis?
The total red cell mass is normal, but plasma volume is reduced
In terms of plasma proteins, how does relative erythrocytosis affect the concentration?
The plasma protein concentration will be increased (not volume!!!!!) as well as PCV, RBC concentration and haemoglobin concentration
What is physiological erythrocytosis?
A mild to moderate temporary increase in PCV may be seen due to splenic contraction which leads to release of erythrocytes into circulation
What is absolute erythrocytosis?
A true increase in total red cell mass, which can be primary or secondary.
What is primary absolute erythrocytosis?
Increased erythrocyte production without increased erythropoietin levels
What is primary absolute erythrocytosis characterised by?
Autonomous erythropoiesis in the presence of low levels of EPO and normal oxygen tension. There is hypoproliferation of bone marrow.
What is Secondary Absolute erythrocytosis?
Increased erythrocyte production in response to increased erythropoietin production
What is appropriate secondary absolute erythrocytosis?
Usually associated with tissue hypoxia, leading to increased EPO production. This stimulates erythrocyte production so that more oxygen can be carried to the tissues.
What is inappropriate secondary absolute erythrocytosis?
Due to increased EPO production without tissue hypoxia. This is a fairly uncommon cause of erythrocytosis.
What is anaemia characterised by?
A reduction in erythrocyte numbers, which results in reduced RBC count, preduced PCV/HCT and usually reduced haemoglobin concentration
What are the clinical signs of anaemia?
Pale mucous membranes, lethargy, reduced exercise tolerance and increased respiratory rate. Heart rate is also increased and there may be murmurs due to turbulence.
Why does anaemia occur?
Decreased production of erythrocytes or increased loss of erythrocytes through destruction or haemorrhage
What is regenerative anaemia?
Where the bone marrow responds to the reduced cell mass by accelerating erythropoiesis and releasing increased numbers of immature erythrocytes into circulation
How can regenerative anaemia be detected?
By the presence of increased number of reticulocytes, anisocytosis (variation in erythrocyte size), polychromasia, basophilic stippling, sometimes macrocytosis and sometimes nucleated erythrocytes.
When is a bone marrow response usually seen following blood loss or RBC destruction?
3-5 days of blood loss
What is non-regenerative anaemia?
Occurs when the bone marrow cannot produce enough erythrocytes to replace those which are lost by normal attrition
Why might non-regenerative anaemia occur?
May be due to deficiency of a factor required for erythrocyte production, inhibition of the bone marrow by certain toxins, infections or neoplasms.
What are pre-regenerative anaemias?
Those in which sufficient time to allow a regenerative response has not elapsed. They will look the same on a CBC as non-regenerative and are differentiated by repeating the CBC 3-5 days later.
In terms of erythrocyte size, what are the 3 classifications of anaemias?
Normocytic: Normal, Macrocytic: Bigger, Microcytic: Smaller
In terms of Haemoglobin concentration, what are the 2 classifications of anaemia?
Normochromic: Normal, Hypochromic: Decreased
What are the two main mechanisms of regenerative anaemia?
Haemorrhage (Erythrocyte loss as whole blood loss) and Haemolysis (Erythrocyte only destruction)
In acute blood loss (haemorrhagic anaemia) what happens to all erythrocyte parameters (PCV, RBC Concentration, Hb Concentration, MCV, ECHC)?
They will be initially normal because the cellular and fluid components are lost in similar proportions.
If blood volume is acutely reduced to 60-70% of normal, what might the animal develop?
Hypovolemic shock
How might the spleen help with PCV count?
Splenic contraction may deliver highly concentrated blood into circulation and temporarily elevate the PCV, but this will be reduced as interstitial fluid attempts to restore blood volume.
In terms of platelets and neutrophils, how might these change during acute blood loss?
Platelet numbers may increase during the first few hours after haemorrhage and mild neutrophilic leukocytosis may occur by ~3 hours.
Describe RBC production following acute blood loss
Increased RBC production in the bone marrow becomes visible in the peripheral blood when reticulocytes are seen. Usually becomes apparent 3-5 days post-haemorrhage
Describe the onset of anaemia and hypovolemia for chronic blood loss
In chronic blood loss, the anaemia develops slowly and hypovolemia does not occur because the animal has time to adapt
Describe the PCV of Chronic Blood Loss
The PCV may reach a low level before clinical signs of anaemia develop
When blood loss is internal, what happens to erythrocytes?
2/3rds of the erythrocytes enter lymphatics and are re-circulated within 24-72 hours. The remainder are lysed or phagocytosed.
When does haemolytic anaemia occur?
When accelerated erythrocyte destruction is not balanced by erythropoiesis.
In terms of plasma protein concentration, what is the difference between haemolytic anaemia and external haemorrhage?
With haemolytic anaemia, the plasma protein concentration is usually normal and in external haemorrhage the plasma protein concentration is usually decreased
When does intravascular haemolysis occur?
Occurs within blood vessels, when the erythrocyte membrane suffers sufficient damage to allow escape of haemoglobin into the plasma
What are some laboratory results characteristic of intravascular haemolysis?
Haemoglobinaemia, Haemoglobinuria, Hyperbilirubinaemia and Alterations in Erythrocyte Morphology
What is Extravascular Haemolysis?
Occurs within cells, and involves accelerated removal of erythrocytes by phagocytic macrophages, especially those in the spleen.
What occurs in immune-mediates haemolytic anaemias?
There is accelerated erythrocyte destruction due to an immunological reaction. The RBCs may be altered antigenically by a drug, infectious agent or be recognised as foreign.
What type of anaemia is usually present in Immune-Mediates Hemolytic Anaemia?
Usually strongly regenerative, however, when antibodies are directed against erythrocyte precursors, anaemia can appear non-regenerative.
What are some haematological features of Immune-Mediated Haemolytic Anaemia?
Marked Regenerative Response, Spherocytosis, Erythrocyte Autoagglutination, Coomb’s Positive Test and Left Shift Neutrophilia
What is neonatal isoerythrolysis?
A form of immune-mediated haemolytic anaemia which occurs in newborn animals
How does neonatal isoerythrolysis arise?
Occurs when a small percentage of horses when a foal inherits from the stallion RBC antigens that are incompatible with those of the mare
Describe the pathogenesis of neonatal isoerythrolysis
Small amounts of foetal RBC antigen gain access to the maternal circulation via foetal-maternal haemorrhage across the placenta. Mare produces iso-antibodies, which enter the colostrum and induce haemolysis of the foal’s RBC’s.
What usually causes iron deficiency?
Almost always due to chronic blood loss, usually into the GIT, but can also occur in suckling animals because of low concentration of iron in milk
Describe Iron Deficiency Anaemia in an acute case vs a chronic case
Iron deficiency is usually regenerative initially, but may become non-regenerative in the late stages
What is the hallmark of Iron Deficiency Anaemia?
Microcytosis - believed to be due to extra cell divisions in erythroid precursors to establish normal intracellular haemoglobin concentrations
Describe a blood smear of a case of severe Iron deficiency anaemia
MCHC is decreased and erythrocytes have an enhanced central pallor, with only a thin rim of peripheral haemoglobin
What are the signs of Sporidesmin Toxicity, and what species does it affect?
It is an acute haemolytic crisis and with haemoglobinuria and jaundice which occurs in adult cattle during the facial eczema season in NZ
How is Leptospirosis Pomona infection characterised in young calves, lambs and weaner deer?
Characterised by an acute haemolytic crisis with anaemia, haemoglobinuria and jaundice
What do Haemolysis produced by L. pomana cause?
Direct lysis of red cells but can also induce immune-mediated haemolysis following adherence to the red cell membrane
What is not a feature of L. Pomana infection in adult animals?
Haemolytic anaemia
What is a feature of Chronic Copper Toxicity in sheep, calves, goats and pigs?
Acute intravascular haemolysis with haemoglobinuria, jaundice and haemoglobinuria nephrosis
Describe the Acute Haemolytic phase of chronic copper toxicity
Occurs when copper is suddenly released from the liver into the bloodstream, usually following some form of stress
Describe how long the period of copper accumulation may be for copper toxicity
May extend from as little as a few weeks to more than a year and will be clinically inapparent
Once the haemolytic crisis of copper toxicity is initiated, what is the sequelae?
Death usually occurs within 2-3 days
What are the two ways by which blood parasites cause Anaemia?
- Immune Mediated Extravascular Haemolysis 2. Intravascular Haemolysis
Describe the immune mediated extravascular haemolysis mechanism caused by blood parasites
Antibodies against the organism bind to the RBC, leading to phagocytosis by macrophage
What are Non-Regenerative Anaemias associated with?
Bone marrow that is unable to produce adequate numbers of erythrocytes, depending on the mechanism neutropenia and thrombocytopenia may also occur
What is Pancytopenia and what is is characteristic of?
Term used when a decrease in all the cell lines is present. Generally associated with Aplastic Anaemia.
When Pancytopenia is present, what does this suggest in terms of cells affected?
Suggests damage to the stem cells, or replacement of the functional haematopoietic tissue with another tissue.
What can cause damage to stem cells?
Drugs, chemicals and toxins, hormones and infectious agents
What are the most common examples of non-regenerative anaemia?
Anaemia of chronic disease and anaemia of chronic renal disease
When might you observe Anaemia of Chronic Disease/Inflammatory Disease?
Can be seen with inflammation/infection, trauma and neoplasia. It is usually mild to moderate, normocytic and normochromic.
What is Anaemia of Chronic Disease/Inflammatory Disease mediated by?
Inflammatory cytokines, and may occur even if there is no clinical or lab evidence of overt inflammation in animal
When might anaemia of chronic renal disease occur?
In chronic renal failure due to decreased erythropoietin production by the kidneys.
Describe the anaemia present in hypothyroidism-related patients
Commonly have a mild normocytic, normochromic, non-regenerative anaemia
Describe the anaemia present in hypoadrenocorticism-related patients
May have mild, normocytic non-regenerative anaemia but can be hard to detect if it is masked by concurrent dehydration
What type of anaemia is Iron Deficiency Anaemia associated with?
Iron deficiency anaemia is usually regenerative initially, and may become non-regenerative in the late stages
Describe the anaemia that can be seen with lead toxicity
A mild non-regenerative or poorly regenerative anaemia may accompany lead toxicity, especially in young dogs
Describe the RBCs and Cells present in Lead Toxicity anaemia
An innapropriately high number of nucleated RBCs and polychromatic cells are present
What is the most characteristic feature of lead toxicity anaemia?
Basophilic stippling of erythrocytes due to the presence of clumped ribosomes. Also inhibits enzymes involved in heme synthesis.
In terms of haemoglobin concentration and size, describe the anaemia of lead toxicity and what is happening in the bone marrow
It is typically normochromic, normocytic with some reticulocytes. There is erythroid hyperplasia and ineffective erythropoiesis in the bone marrow
What are the 5 focus points used to assess RBC in a blood smear?
Number, Size, Shape, Colour and ‘Other’ - ‘Other’ refers to structures in or on the erythrocytes and unusualy arrangements on the blood film
How is erythrocyte number measured?
By the RBC indices in the CBC or by measuing a PCV. Cannot be estimated accurately from a blood smear alone.
What is Anisocytosis?
The variation in RBC cell size, can be due to the presence of large cells, small cells or both
What is Microcytosis?
Presence of smaller erythrocytes
What is Macrocytosis?
Presence of large red blood cells
What are the hallmarks of regenerative anaemia on a blood smear?
Anisocytosis and Polychromasia
What is hypochromasia?
Refers to increased central pallor of cells due to decreased haemoglobin content - i.e. less colour
What is Polychromasia?
Refers to the variation in colour of erythrocytes, caused by the presence of more basophilic reticulocytes in addition to normal mature erythrocytes
What are Polychromatophils?
Immature/younger erythrocytes that have been released earlier from the bone marrow
What are reticculocytes?
Large, non-nucleated erythrocyte containing sufficient RNA/organelles to stain as granules
What dose an increased number of reticulocytes indicate?
Presence of regenerative anaemia in most species
What is the general term for any abnormally shaped erythrocytes?
Poikilocytosis
What are Acanthocytes?
Cells with few too many irregular surface projections of varying length.
What causes Acanthocytes?
Thought to occur due to changes in cholesterol/phospholipid concentrations in the red cell membrane
What are Echinocytes?
These have multiple, short, blunt, evenly spaced surface projections and are most commonly seen due to drying of a blood smear
What are Schistocytes?
Irregularly shaped erythrocyte fragments due to intravascular trauma
What are Spherocytes?
Small, dark staining, spherical erythrocytes which result from partial phagocytes by macrophages in the spleen and liver
What are Eccentrocytes?
Cells that have their Hb shifted to one side due to oxidative damage. The end result is a clear area to the side of the cell
What is meant by Basophilic Stippling?
Refers to erythrocytes with blue dots in the cytoplasm. Occurs in ruminants and cats as part of regenerative anaemia.
If basophilic stippling is seen without anaemia and reticulocytosis, what is it indicative of?
Lead poisoning
What are Heinz Bodies?
Clumps of denatured haemoglobin on the internal surface of the erythrocyte membrane due to oxidative damage. Most cats have these in up to 10% of their erythrocytes
What are Howell-Jolly bodies?
Nuclear remnants. They appear as small, darkly staining, spherical bodies within erythrocytes. Usually occur with regenerative anaemia
What are Metarubicytes?
They are the last nucleated stage of erythrocytes and can be seen in several stages including the regenerative response and bone marrow damage.
What are Siderocytes?
These mature erythrocytes contain iron granules and are seen with lead poisoning and certain anaemias
What is useful about a Bone Marrow Examination?
Can help to confirm or exclude differential daignoses based on the examination of peripheral blood and provide a more reliable indciation of prognosis.
What are the indications of Bone Marrow Examination?
Non-regenerative Anaemia, Persistent Leucopenia, Thrombocytopenia and Presence of Atypical Cells Suggestive of Neoplasia
What should be collected in addition to a bone marrow aspirate, during the bone marrow examination?
Always collect a peripheral blood sample for a CBC at the same time, as bone marrow aspirates need to be interpreted with concurrent peripheral blood results.
What is the function of Leukocytes?
They are primarily involved in body defence mechanisms and are capable of countering a wide variety of different infectious agents or foreign material
What are the 2 basic mechanisms for which granulocytes defeat the baddies?
Phagocytosis and Antibody production
What leukocytes have phagocytic properties?
Granulocytes and Monocytes
What cells are grouped within ‘Granulocytes’?
Neutrophils, eosinophils and basophils