HLI 3 Flashcards
(93 cards)
1
Q
Where are blood cells produced
A
Early embryo – yolk sac
Foetus – liver, spleen and bone marrow
Neonate – liver and bone marrow
Adult – bone marrow (spleen & liver with disease and when needed - EMH)
2
Q
What is leukopoesis divided into and how long do they live
A
- Lymphopoiesis
- Myelopoiesis
- Granulocytes
- Neutrophils
- Eosinophils
- Basophils
- Monocytes leads to macrophages
Most have a life span of 2 weeks
3
Q
granule colour of neutrophil, basophil and eosinophils
A
Neutrophils - granules cannot see
Basophils - granules blue
Eosinophils - granules red
4
Q
Why do leukocytes undergo segmentation
A
Elongated segmented nucleus
- Easier to get through endothelial cells into the tissues - first to arrive
- Inactive nucleus no transcription
5
Q
Where are neutrophils found in the body and how long in each area
A
- Marrow
○ Maturation pool
○ Storage pool - dogs and cats store up to 5 days - Blood
○ Circulating neutrophil pool (free moving in vessels) - what we get in the blood sample
○ Marginal neutrophil pool (loosely adhered to vessels)
- 10-12 hours - Tissue - 24-48 hours
- do not recirculate unlike lymphocytes (removal via macrophages)
6
Q
Monocyte kinetics
A
- Maturation in marrow is rapid (24-36hrs)
- Don’t have to have the segmented nucleus or protein granules produced
Lifespan - unknown
Limited recirculating and replication capacity
Continue to divide
7
Q
Thrombopoeisis what regulated by, timeframe
A
- Regulated by thrombopoeitin from liver, kidney and marrow stromal cells
- Maturation 2-10 days
8
Q
What is a megakaryocyte and what does it lead to
also what are macro platelets and their implications
A
Megakaryocyte
- Large mass of nuclear material
- Can be 34 times the normal amount of DNA
- Cytoplasm pinching off that become the platelets
Macro platelets or macro thrombocytes
- Produced from larger pinching of the megakaryocyte when need lots of platelets
- If platelet larger than RBC then classify as this
9
Q
What are the 4 general mechanisms for decrease platelets and examples
A
1) production - bone marrow disease or EMH
2) consumption - inflammation
3) sequestration - splenomegaly from haemangiosarcoma
4) destruction - immune mediated
10
Q
What percentage of platelets are sequestered in the spleen, life-span and when nucleated
A
- 30-40% sequestered in spleen within vascular sinusoids, if get splenomegaly then holds more
- Life-span 5 – 9 days
- Nucleated in avians and reptiles (thrombocytes)
11
Q
What are the 3 features of the leukogram
A
- Total leukocyte count
- Differential (individual leukocyte counts)
- Leukocyte morphology - human only,
12
Q
White blood cell count what are the 3 ways
A
- Automated count methods
◦ Impedance
◦ Flow cytometry - Manual count
◦ Evaluate in monolayer
◦ Est. WBC count x 109/L = 2.0 x average WBC per 40x field - Assess morphology (40 or 100x objective in monolayer)
13
Q
What are the 3 features of the leukogram
A
- Total leukocyte count
- Differential (individual leukocyte counts)
- Leukocyte morphology - human only,
14
Q
White blood cell count what are the 3 ways
A
- Automated count methods
◦ Impedance
◦ Flow cytometry - Manual count
◦ Evaluate in monolayer
◦ Est. WBC count x 109/L = 2.0 x average WBC per 40x field - Assess morphology (40 or 100x objective in monolayer)
15
Q
What are the 3 characterstics that automated systems determine in WBC and when should and shouldn’t it be used
A
- Cell size - forward scatter
- Cell fluorescence - RNA within the cell
- Side scatter (cell complexity) - granularity
Good with normal leukocytes but misclassifies cells that are abnormal - look at blood smear
16
Q
Neutrophil kinetics for cows, dogs, horses and cats
A
Cows - 1:1 easily become neutropenic due to small storage pool
Dogs - 1:1 large storage pool (5 days worth)
Horses - 1:1 small storage pool not as small as cows
Cats - 3:1 most neutrophils in the marginating pool, smaller storage pool than dogs but still larger than horse and cow
17
Q
what occurs to neutrophils during the stress response
A
SUPPRESS IMMUNE SYSTEM
lymphocytes release from bone marrow is decreased
movement of RBC from marginating into circulating pool
- mature neutrophilia
Move red blood cells from marginating into circulating
Neutrophils are also released from the bone marrow
18
Q
What occurs to neutrophils during inflammation
A
neutrophilia - degree varies
pull from storage pool into circulating and marginating +/- increased marrow production
19
Q
what occurs to neutrophils during excitement (adrenalin mediated)
A
Mainly in young animals
neutrophilia
- increase in lymphocytes and neutrophils movement from marginating into circulating pool
20
Q
List 4 causes of neutropenia and how occurs
A
- Overwhelming inflammatory demand
- Generally a septic process with a lot of puss formed - pyometra
- Less disease needed for cows as they have a lower storage pool - Transient margination (endotoxaemia)
- More towards edges of the vessels so not detecting them in the sample - Bone marrow disease
- Make less neutrophils - Immune-mediated destruction of neutrophils
21
Q
what results in a neutrophil left shift and the types
A
= immature neutrophils in blood
• Usually bands
• Regenerative left shift
○ ◦ Mature (segmented) neutrophils > immature (band) neutrophils
• Degenerative left shift
○ ◦ Immature neutrophils > mature nature neutrophils
Can have with normal neurophils
22
Q
What does a degenerative left shift indicate
A
Tells us bone marrow is desperate - no longer have mature left
More likely to be septic and life threatening - severe inflammation
23
Q
List 2 causes of neutrophil left shift
A
- Inflammation
= High tissue demand for neutrophils
- Reflects release from maturation pool - Can also occur with myeloproliferative disease e.g. chronic myeloid leukaemia (serious bone marrow disease) and neoplasia (paraneoplastic syndrome)
24
Q
List 4 differentials that lead to lymphocytosis
A
1. Chronic antigenic stimulation ◦ Vaccination 2. Adrenaline/Excitement (young animals) 3. Lymphoid neoplasia 4. Hypoadrenocorticism - lack of cortisol
25
List 3 differentials that lead to lymphopenia
```
1. Glucocorticoids/Stress - most common
◦ Reduced release from LN and spleen
◦ Lympholysis
2. Acute inflammation
- Generally increased stress so get lympholysis or due to movement of lymphocytes into tissues - less circulating
3. Loss of lymphatic fluid
◦ Chylothorax
◦ Enteric neoplasia
◦ Protein losing enteropathy
```
26
List characteristics of monocytes
1) variable nuclear morphology
2) blue-grey cytoplasm
3) larger than neutrophils
4) cytoplasmic vacuoles
5) nuclear never as dense as neutrophils
27
List function of monocytes
1) phagocytosis - foreign material, daed cells
2) source of cytokine and chemotactic factors
3) present antigen to T cells
4) perform antibody-dependent cytotoxicity
28
List 4 differentials that lead to monocytosis and what does mnocytopenia mean
1. Acute inflammation
2. Chronic inflammation
3. Glucocorticoids/stress (dogs)
4. Myeloproliferative disease e.g. myeloid leukaemia
clinically insignificant - mainly healthy animals have low numbers
29
List 3 differentials for eosinopenia
1. Glucocorticoids/stress response
| 2. Can be clinically insignificant
30
What are the 4 functions of basophils and what are they associated with
1. Histamine release
2. Promote lipid metabolism
3. Haemostasis
4. Parasite control
Generally occur with eosinophils
- Eosinophilia generally get basophilia
31
How does a scatter plot work for WBC differentiation
each cell going through the analyser gets placed on graph based on fluorescence and granularity - so each type of leukocyte will be placed in a certain area - look at how many dots in that area to see the amount of neutrophils, basophils etc.
32
How does a scatter plot work for WBC differentiation
each cell going through the analyser gets placed on graph based on fluorescence and granularity - so each type of leukocyte will be placed in a certain area - look at how many dots in that area to see the amount of neutrophils, basophils etc
- Analyser needs to know the species as the cells change characteristics in different species so certain cells may be identified as something they are not
33
What is toxic change, Why and when does it occur
What?
- Neutrophils that show signs of cytoplasmic immaturity
- Was able to divide the nucleus but cytoplasm wasn't divided properly
Why?
- Hastened or disordered maturation in the bone marrow
When?
- Infections or intense inflammation or myeloproliferative disease
Can be associated with nuclear immaturity (left shifting) or seen in cells with mature segmented nuclei
34
What are the 3 main signs of toxic change
1) increase basophilia
2) dolhe bodies
3) vacuoles
35
Ring form neutrophils when seen
low numbers in normal rodents
◦ intense inflammation
◦ chronic myeloid leukaemia
◦ myelodysplasia.
36
hypersegmented neutrophils how many segments considered and causes
- >5 segmentations = Older neutrophils
- Chronic glucocorticoid exposure - MOST COMMON
○ decreased tissue emigration - less margination more circulating neutrophils - age in the blood
- Seen more in horses - normally have some with higher segmentation - 6
- Can also be seen with myeloproliferative disease e.g. chronic granulocytic leukaemia
37
Describe botyroid nucleus and barr body
Botryoid nucleus - hyperthermia - heat stroke
- Cooking of the neutrophils
- Can be high fever
Barr body
- Only found in females as need two X chromosomes
- One X chromosome is inactivated and forms the barr body
- Present in all female neutrophils but won't see every time due to orientation of nucleus
38
Describe reactive lymphocytes and what are they hard to differentiate between
Larger than normal
- (nucleus 2x rbc)
Increased basophilic cytoplasm
- May have few magenta cytoplasmic granules
- Reflect increased antigenic stimulation
- Non-specific, often seen in health
Hard to differentiate between REACTIVE and NEOPLASTIC
39
What are 2 types of cytoplasmic inclusions and conditions that lead to them
```
1, erythrophagia
2. sideroleukocyte
What conditions might lead to these cells in the blood?
1.. Previous blood transfusion
2. IMHA
```
40
What occurs with Pelger-Huet Anomaly
failure of nuclear segmentation
- lots of band and no segmental neutrophils - weird
- still functional and NO TOXIC CHANGE
41
What characteristics define an acute inflammatory leukogram and why
Neutrophilia*
- Sometimes don't have neutrophilia yet - snapshot in time
+/- Toxic change - most important
+/- Left shift
+/- Monocytosis
*most common indicator but not always present
Neutrophilia with Inflammation
= Release of neutrophils from storage pool +/- release from maturation pool (bands) + increased production (with time)
42
What disease might cause an acute inflammatory leukogram
```
A focus of suppurative inflammation or sepsis e.g.
Not heaps of puss
- Infected wound
- Pneumonia
- Pyelonephritis
- Tissue damage and necrosis
```
43
List some species variation in response to acute inflammation: cat and dogs, cattle and horses
1) Cats and dogs often have WBC 10-30 x 109/L with inflammation
○ Due to the large storage pool
2) Cattle may only show hyperfibrinogenaemia with no inflammatory leukogram with acute infections
○ Cattle often show transient leukopenia (small storage pool)
3) Horses have moderate leukocytosis and left shift response, often WBC 7-20 x 109/L with acute inflammation
44
At what level of neutrophilia do you start getting worries about leukaemia
50-100 x 109/L
| ○ Above 50 start getting worried about leukaemia
45
What characteristcs define a severe acute inflammatory leukogram
“Leukaemoid response”
- Marked Neutrophilia - 50-100 x 109/L
○ Above 50 start getting worried about leukaemia
- Regenerative left shift - pooling out mature cells
○ Bands - less
○ Metamyelocytes - even less
○ +/- Myelocytes
46
What diseases might cause a severe acute inflammatory leukogram
A focus of intense suppurative inflammation or sepsis
- Pneumonia
- Pyothorax
- Peritonitis
- Pyometron
- Necrotic tumour (within tumour)
○ DDx: Myeloid Leukaemia, Paraneoplastic syndrome - produce cytokines - increase inflammation
47
what characteristics define overwhelming inflammatory leukogram
Neutropenia and or degenerative left shift (not putting out mainly mature now also immature)
- +/- Lymphopenia - don't leave lymph node as quickly
- +/- Monocytosis
48
What diseases might cause an overwhelming inflammatory leukogram and what prognosis
A focus of acute sepsis e.g. - pouring pus
1. Intestinal perforation - gut rupture
2. Ruptured Pyometron
3. Endotoxaemia, Gram –ve infections
= Marrow unable to meet demand - potential for animal to die is higher
DDx: Bone Marrow Disease
49
What characteristics define chronic inflammatory leukogram
- Neutrophilia - mature
- +/- Slight regenerative left shift
- +/- Lymphocytosis
- Monocytosis (high)
50
what diseases might cause chronic inflammatory leukogram
A focus of chronic inflammation e.g.
1. Severe pyoderma
2. Necrotic neoplasia
3. Chronic hepatitis
4. Fungal infections
51
What are the characteristics of a glycocorticoid/stress leukogram and the reason for each
```
Lymphopenia
◦Decreased release from nodes
◦Margination
Neutrophilia (Heterophilia in birds)
◦Increased release from storage pool
◦Shift from marginating to circulating pool (express less adhesion factors)
Eosinopenia
◦Decreased release from marrow
Monocytosis (dogs)
◦Shift from marginating to circulating pool
NO LEFT SHIFT OR TOXIC CHANGE
```
52
list the magnitude for the stress leukogram in cats, dogs, equine, bovine
○ Feline 3:1
○ Canine, equine, bovine 1:1
- Expect mature neutrophilia up to 3x neutrophil upper reference interval in cats
- Expect mature neutrophilia up to 2x neutrophil upper reference interval in dogs, horses, cows
53
what is a physiologic leukocytosis and what characteristics define it
```
= Adrenalin/excitement mediated - short duration and generally younger animals
- Neutrophilia
- Lymphocytosis
- +/- Monocytosis
- Young animals
NO left shift or toxic change
```
54
why do you see a neutrophilia, lymphoctytosis
Neutrophilia
•Shift marginating to circulating pool (Increased blood pressure and HR) - very temporally - take two time apart and may go back to normal
Lymphocytosis
- Shift marginating to circulating pool
55
Granulocytic hypoplasia leukogram what is it, what is it caused by
Persistent Neutropenia with no left shift (>5 days) - bone marrow takes 4-6 days to produce
- Caused by bone marrow production problem e.g.
◦Immune mediated neutropenia
◦Drug toxicity - chemotherapy
56
How long should we wait before doing bone marrow evaluation on a neutropenic patient?
5-7 days
57
what is the point at which neutropenia is dangerous
once get below 1.5x10^9 put on antibiotics as risk of getting infection and not being able to fight
58
Hypoadrenocorticism Leukogram (Addisons syndrome) what characteristics define and the electrolyte
- Lack of stress leukogram despite chronic or severe illness - consider this in dogs
○ Could also be excitement - more likely in cats
- +/- Lymphocytosis-
- +/- Eosinophilia
- Hyperkalaemia
- Hyponatraemia and hypochloridaemia
- Low Na: K ratio (<27:1)
59
where are most plasma proteins produced and which ones aren't
synthesised in the liver
| - immunoglobulins are produced by B cells
60
What are the main functions of plasma proteins
```
• Exert colloidal osmotic pressure
– maintains intravascular fluid volume
• Antibodies (immunoglobulin)
• Coagulation factors
• Hormones
• Enzymes
• Help maintain acid-base balance
• Transportation of substances
```
61
What are the 2 types of plasma proteins, how much do they compromise of and examples
1) albudmin - 55% of total protein, accounts for 75% of colloid osmotic activity
2) globulins - diverse group divided into 3 main groups
1. alpha
2. beta - complement, IgM, IgA
3. gamma, IgG, C-reactive protein
62
What are the 3 main ways to measure plasma proteins and what do they measure
1. Using plasma via refractometer
– Total Solids (an estimate of total protein)
2. Using serum via laboratory analyser
– Total Protein + albumin
3. By serum protein electrophoresis - uncommon
63
Refractometer what measures in terms of protein, what tube used, what is the issue and what else does it measure
- using plasma (from whole blood) - use EDTA tube
- measured total solids
- false increase can be due to glucose, urea, sodium, chloride, lipaemia
- fibrinogen is present - used to estimate fibrinogen - heat preipitation to remove solids and leave fibrinogen
64
biochemistry analyser what does it measure and what else can you measure
- measures total protein and albumin - so learn more about proteins
globulins are then calculated [Globulins] = [Total Protein] – [Albumin]
65
serum protein electrophoresis (SPE) what is involved and how used
nout a routine test but allows quantification and separation of globulins
- patterns help with differential diagnoses
66
What are the two types of abnormal protein concetrations
1. Selective
• one is changed in a different direction than the other
eg glomerular disease (albumin is lost – hypoalbuminemia)
2. Non-selective
• both change in the same direction - hypoalbuminemia and hypoglobulinaemia
• A/G ratio remains unchanged
eg haemorrhage (both lost – panhypoproteinemia)
67
Give a cause of increased TS and decrease TS
Increased TS = hyperproteinaemia
- dehydration
Decreased TS = hypoproteinaemia
- haemorrhage
68
List 3 reasons for hyperglobulinaemia
1) Dehydration
– relative increase due to plasma water loss
2) Inflammation
– absolute increase due to production of positive acute phase proteins and immunoglobulins
3) Less commonly see with B-lymphocyte neoplasia - use serum protein electrophoresis to determine
eg multiple myeloma or less commonly B-cell lymphoma
69
What are the characteristics in a serum protein electrophoresis for inflammatory disease and neoplasia
inflammatory
- polyclonal gammopathy = wide-based immunoglobulin peak on electrophoresis - more than one cell line increased
neoplasa
- monoclonal gammopathy = narrow-based immunoglobulin peak on electrophoresis
70
What are the 3 main mechanism for hypoalbuminaemia and give 2 examples
1) increased albumin loss
- haemorrhage, gastrointestinal.renal loss
2) decreased albumin production
- hepatic insufficiency, inflammation, malabsorption
3) haemodilution - uncommon
71
List clinical effects of hypoalbuminaemia and at what concentration does that occur
```
When plasma albumin loss is marked ie < 15 g/L:
• Colloid osmotic pressure decreases
• Can lead to vascular fluid loss into tissues:
- Peritoneal effusion (ascites)
- Pleural effusion
- Pericardial effusion
- Pulmonary oedema
- Submandibular oedema
```
72
What are the 4 main mechanisms of hypoglobulinaemia and examples
1. • Increased loss
- haemorrhage, GIT, skin, effusions
2. • Decreased production
- malabsorption, maldigestion, malnutrition
3. • Failure of passive transfer
- inadequate colostrum intake in the newborn - measure immunoglobulins
• horses and cattle
4. • Severe combined immunodeficiency syndrome (SCID) - rare
- Horses such as arabs are pre-disposed as well as some dog breeds - jack Russell
73
List 4 positive acute phase proteins
1) fibrinogen - moderate APP, rises slowly, readily available
2) haptoglobin (cattle)
3) serum amyloid A
4) C-reactive protein - dogs
Bottom 3 are major and rapidly rise and also rapidly decrease resulting in them being very sensitive
74
List the main negative APP ad what is the APP testing used for
- decreases with inflammation
Albumin
- can take several days to decrease due to longer half-life
Testing
- monitor for post-operative infection - cheap way, should follow normal increase-decrease pattern
75
What are the 2 main causes of hyperfibrinogenaemia
```
1. Inflammation
• positive acute phase protein
§ useful in horses and cattle
§ may increase before leukogram changes
2. Dehydration
• relative increase due to plasma water loss
```
76
what are the 2 main causes for hypofibrinogenaemia
1. Decreased synthesis
- reduced functional hepatic mass
• more than 70-80% loss of function
2. Increased consumption
- intravascular coagulation ie DIC
77
when are you clinical suspicious of bone marrow disease
haematologic abnormalities are present persistent and not readily explained
such as 1. decrease in blood cell line or increase, atypical immature cells, hypercalcaemia
78
when are you clinical suspicious of bone marrow disease and list the 6 abnormalities
haematologic abnormalities are present persistent and not readily explained
1. Decrease in a blood cell line
2. Severe increase in a blood cell line
3. Atypical or immature cells
4. Marked hyperproteinemia
5. Hypercalcaemia
79
What is the most common indicators of bone marrow disease and give 3 examples
Decreased cell numbers
can be one, two or all three - pancytopenia
1. persistent non-regenerative anemia - no polychromasia
2. persistent neutropaenia - no left shift or toxic change
3. thrombocytopaenia - no large or giant platelets, run coag panel to rule out DIC
80
List 5 causes of decreased cell numbers
1) infectious - parvovirus, FeLV
2) toxins - bracken fern - ruminants
3) immune-mediated disease - IMHA
4) endocrine disease - hypothyroidism
5) neoplasia - replace bone marrow
81
List 3 examples of what cause severe increase in cell numbers
1. Erythrocytosis, rubricytosis
• no evidence of splenic contraction, dehydration, hypoxia, renal disease
2. Leukocytosis
• no evidence of inflammation
3. Thrombocytosis
• no evidence of Fe deficiency, inflammation
82
Differences between leukaemia and lymphoma
1. Leukaemia
• neoplastic haematopoietic cells originate in bone marrow but often seen in circulation
○ Could be red blood cell, platelet, neutrophil
○ acute vs chronic
○ lymphoid vs myeloid (all cell lines except lymphoid)
2. Lymphoma
• neoplastic lymphocytes originate in solid tissue
lymphoid tissue outside the bone marrow eg lymph node, spleen, liver, intestine, skin
83
How to differentiate between lymphoma and leukaemia
Look for original site of lymphoma - lymph nodes huge!
Happy dog as lymph nodes not painful
Chemotherapy can increase quality of life for 1-2years
Leukaemia dog - very sick, dead within a couple of weeks
84
What are 2 types of atypical cells and what do they indicate
indicate bone marrow disease
1. immature cells - abscence of regnerative response
2. abnormal cell
85
Marked hyperproteinaemia what does it support bone marrow evaluation for
1. Lymphoid neoplasia
• B-cell chronic lymphocytic lymphoma
• Multiple myeloma (plasma cell neoplasia)
2. Systemic fungal and protozoal infections - can see in bone marrow aspirates
• Histoplasmosis, Leishmaniasis
○ both exotic to Australian domestic animals
86
hypercalcemia what 3 main neoplasms lead to this
1. lymphoid neoplasms
- production of PTH-related protein
2. multiple myeloma
- localised osteolysis
3. metastatic neoplasia (eg histiocytic sarcoma)
- production of PTH-rp or VitD
87
what are the two main bone marrow collection and what are they good for
Simultaneous evaluation of bone marrow cytology (aspirates - great for cell morphology)) and histology (core biopsy - evaluate architecture) is recommended
88
What are the 4 site selections for bone marrow
```
1. Pelvis
• iliac crest; medium/large size dogs
2. Proximal femur
• trochanteric fossa; cats and small dogs
3. Proximal humerus - go to site
• common site especially in obese animals
4. Sternum
• site of choice for horses
```
89
When do you need to make the smears for bone marrow collection and what must you submit
- smears are made immediately after collection
- submit several unstained smears
- must be submitted with an EDTA blood sample within 24 hours
90
multiple myeloma what type of neoplasm and what characteristics present
- plasma cell neoplasia - malignant - specialised B-lymphocyte
Diagnostic
1. neoplastic plasma cell in bone marrow
2. monoclonal gammopathy
3) oesteolytic lesions
4) light chain proetinuria
91
List the lymph nodes associated area of drainage
1) Axillary lymph node
2) Mediastinal lymph node
3) Mesenteric lymph node - intestine
4) Popliteal lymph node
5) Prescapular lymph nodes
6) Retrophoryngeal lymph node
7) Submandibular lymph node
1) distal forelimb
2) lungs
3) intestine
4) distal hind limb
5) shoulder
6) pharynx
7) mandible
92
what is different about total calcium in the blood and what effects this
70% bound to albumin (not active form) so if get increase in albumin will get increase in calcium
If albumin is high due to many reasons such as dehydration
93
What interferes with ADH
- Lack of ADH production or issue with kidneys response to ADH (determine final concentration of urine - can draw in huge amount of water)
- Corticosteroids, toxins released by bacterial infections (pyometra), hypercalcaemia
- This can be due to hypercalcaemia - also causes alterations in renal blood flow - decreased filtration rate, renal mineralisation overtime - leads to full renal problems
