Midterm 1 (1-7) Flashcards
How to determine BT, BMBT
Use a sharp, sterile blade, make 0.1-0.2mm deep, 0.5cm long incision on inner ear skin. Wipe the blood flowing from under the wound in 20-30 second intervals. Measure time from appearance of first drop till the end of bleeding.
Normal BMBT- 3-5mins
for vasculopathies, thrombocytopathies and thrombocytopenias
What is normal BT?
3-5 mins
What is DIC
Disseminated intravascular coagulopathy
common acute disorder (+FDP or D-dimer= sign)
Fibrinolysis and microthrombus function are present in diff parts of the body simultaneously due to severe tissue damage and blood vessel injury- initiation of intrinsic pathway (coag factors and platelets- consumpted quickly)
Lab signs of DIC
increased coagulation time, bleeding time, PT, APTT, TT, FDP
decreased platelet count
appearance of schysocytes and/ or Burr cells in blood smear
What is thrombocytopathy?
Decreased ability of platelets to aggregate and adhere to the site of injury- forms primary thrombocyte-thrombus
Causes of thrombocytopathy?
Improper development of platelets, von Willebrand disease (doberman), uremia, liver failure, NSAID treatment, myelo- or/ and lympho-proliferative disease.
Regular sizes of thrombocytes
1-2um, centre is granulomer and edge is hyamoler
eq, bo, sheep= 3-5fl
ca, su= 7-8fl
fe= 10-15fl
Causes of thrombocytopenia
Causes-
decreased production in bone marrow
increased utilisation: DIC
increased loss, destruction (AITP), sequestration (chronic splenomegaly)
ESR
Erythrocyte sedimentation rate- increased sedimentation is due to inflammatory processes (also checked with glutaric aldehyde test). Increased sedimentation means that the APP and other globulins attach to the surface of the RBC.
It is faster in eq, so must evaluate after 20 mins (decreases due to inflammation, opposite of other species).
ESR is inversely proportional with Ht
ESR is directly proportional with viscosity, total protein and fibrinogen concentration.
Causes of hyperlipidemia
increased fat content in diet, DM, hypothyroidism, hyperadrenocorticism, nephrotic syndrome, septicemia, pancreatitis.
Causes of hypolipidemia
starvation (long term), chronic liver failure
Causes of hypercholesterolemia
Increased dietary fat content, hypothyroidism, hyperadrenocorticism, nephrotic syndrome, DM
causes of hypocholesterolemia
liver failure, neoplastic disease, hyperthyreosis, malnutrition and malabsorption
What is the total cholesterol and cholesterol ester?
2-6mmol/l (ester is 40% of this)
Causes of hyperglycemia?
Transient- Lab errors, stress (fe), food intake (ca, humans), xylazine effect, cranial trauma/inflammation (Aujeskzy disease, rabies), after/during glucose fluid therapy
Constant- DM, hyperadrenocorticism, progesterone effect, enterotaxemia (sheep)
Causes of hypoglycaemia?
Lab error, decreased energy (ketosis in Ru, growing pigs, racing, starvation), insulin overdose, liver failure, hypoadrenocorticism, septicaemia, hyperthyroidism
What is monoclonal gammopathy?
one protein fraction derived from one clone- immune mediated or neoplastic conditions
What is polyclonal gammopathy?
beta and gamma globulins derived from different clones- inflammatory processes or some immune mediated diseases
Hyperglobulinemia
polyclonal gammopathy- broad peak in gamma and beta region
common causes- chronic inflammation, liver disease, FIP, occult heartworm disease, Ehrlichiosis
Beta- gamma bridging- disorders with increased IgA and IgM- heartworm, lymphoma, chronic active hepatitis
Monoclonal gammopathy- sharp spike in beta/gamma region, neoplastic or non-neoplastic
Neoplasia- common cause- myelomas
other causes- chronic lymphocytic leukemia (IgG), lymphoma (IgM, IgG), extramedullary plasmacytomas (solid tumor with plasma cells) in GI tract, skin and liver (Car)
Increase of IgM= macroglobulinemia (Waldenstrom, neoplasm of B-cells accompanied by splenomegaly or/and hepatomegaly, but lacking osteolytic lesions)
Multiple myelomas- disorder of plasma cells that have undergone antigenic stimulation in peripheral lymph nodes, with osteolytic lesions)
Non neoplasmic (rare)- occult heartworm, FIPV, Ehrlichia canis, lymphoplasmacytic enteritis and dermatitis, amyloidosis.
Hypoglobulinemia
decreased intake (in neonates before drinking colostrum, absorption disorders) decreased synthesis- liver failure, or inherited immunodeficiency increased loss- PLE, PLN, skin (burning, inflammation), bleeding
Reticulocytes
young but differenciated RBC with basophil punctuates stained by brylliant-cresil blue (big blue= younger), have same function, can carry O2.
Not in eq, Ru (only in bone marrow)
Appearance is sign of regenerative function of bone marrow.
If they’re nucleated, they’re too young and can’t carry O2.
In case of maturation arrest (vit B12, folic acid deficiency or feline leukemia) they won’t turn into reticulocytes.
Fresh EDTA blood and Brylliant cresil (in same proportion), physiological saline and 3.8% Na-citrate and mix. Incubate 2-3 hours, prepare smear.
2-3% reticulocytes per 100-1000 RBCs.
CRC, CRP
Increased count- acute blood loss (3-5 days needed for bone marrow to increase reticulocyte count), haemolytic anema, chronic blood loss, some nutrient deficiency anemia.
Staining for reticulocytes
.04g Brylliant-cresol blue+ 8ml physiological saline solution. Mix
Add 2ml 3.8% Na-citrate and mix.
Mix equal amount of fresh EDTA blood
Make blood smear
Normal number of reticulocytes?
In 100-1000 RBC, 2-3% are reticulocytes
Typical changes in derived parameters? MCHC, MCV, MCH etc
macrocytic, hypochromic (increased MCV, decreased MCHC) with increased reticulocytes= regenerative anemia
normocytic, normochromic (normal MCV and MCHC, possible decreased MCH)= non regenerative
microcytic, hypochromic (decreased MCV and MCHC)= iron, copper, pyridoxine deficiency anemias, liver failure, portosystemic shunt (with normochromic- normal for Akita)
macrocytic, normochromic (impaired DNA synthesis)- Bit12, cobalt or folic acid deficiency, erythroleukemia, poodle macrocytosis)
RDW of dog an cat?
Red Cell Distribution
Dog- 12-16%
Cat 14-18%
What does short RDW mean?
Non regenerative processes
Normochromic value (MCHC)
300-350g/l
Hyperchromasia effects?
erythroleukemia
vitamin B12, cobalt or folic acid deficiency
splenectomy
lead poisoning
Hypochromasia effects?
iron, copper, pyridoxine deficiency anemias, newborn animals, liver failure
How to measure Hbg?
Spectrophotometer method (drabkin method)
Describe spectrophotometer method for Hgb measurement
Put 20ul whole blood sample to 5ml reagent (K3SE(SCN))6 haemolyses RBC and forms Fe3+ from Fe2+, which is further oxidised by KCN to cianidmethaemoglobin.
Mix and measure amount of orange end product.
Normal Hbg amount?
18-20mmol/l
How does O2 binding capacity of Hgb increase?
decreased 2,3-DPG levels in RBC, CO2 levels (resp alkalosis), temperature (hypothermia) increased pH (met/resp alkalosis)
How does O2 binding capacity of Hbg decrease?
Increased 2,3-DPG level of RBCs, Co2 (resp acidosis), temp (hyperthermia) Decreased pH (met/resp acidosis)
When does Hgb conc. increase?
relative (dehydration) or absolute polycythemia
When does Hgb conc. decrease?
Relative (hyperhydration) or absolute anema
TWC, EC, IC
600-650ml/kgbw
250-300ml/kgbw
350-400ml/kgbw
Blood clotting parameters
Na2-citrate as anticoagulant- suitable for blood smears and binds calcium reversibly
Citrate blood ratio for homeostasis?
1:9 (but in RBC sedimentation test, 1:4)
Blood biochemistry parameters measured in…?
Serum, without anticoagulant or blood plasma with heparin (anticoagulant)
Volume disturbances?
Perfusion and hydration disorders, PCV and Ht (increase for dehydration), Hb conc (increases for dehydration), TP/Talb conc (increase for dehydration), MCV or RBCs
Evaluation of perfusion (IV deficit and circulation problems)
CRT (hypo/hypervolemia)
color of mucous membranes (pale, livid)
strength of pulse
heart rate (increased BP)
Evaluation of hydration
skin and eye turgor
mucous membranes (wet, shiny, dry etc)
skin around oral/anal cavity (signs of water loss)
sunken eyes (enophthalmos), prolapse of 3rd eyelid (fe)
changing of weight
urine volume
Examining blood in Ht tubes after centrifuging
- plasma color change-
usually transparent (eq and bo yellow due to bilirubin and carotenoid)
dark yellow- hyperbilirubinemia
red- haemolysis
white- hyperlipidemia
chocolate (methaemoglobinema) - buffy coat (width changes w/ WBC count)
- microfilaria larvae- on top of buffy coat, sometimes visible
Volume loss in acute bleeding?
5-15% no change in BP
15-25% vasoconstriction, initial increase in BP, tachycardia
35-45%- severe decrease in BP, oliguria/anuria, vasodilation leading to shock
50%- death
PCV methods
- microcapillary- homogenous and fill almost full, plug one end with cold plasticine and centrifuge 3-5min
- automated cell counter- MCVxRBC/1000
- Handheld HCT meter- Ht and Hgb in whole blood. species chip and test strip into meter and add drop of whole blood to test strip.
Normocytic, hypovolemia?
during acute blood loss or immediately after- shock!
normocytic, hypervolemia?
overdosing on blood transfusion/chronic stress (high BP)
Polycythemia, normovolemia
- False- long time in EDTA, swelling
- Physiological- congenital, newborn or long term hypoxia
- Absolute-
increase in RBC
primary (w/o EPO)- bone marrow neoplasia, chronic leukemia
secondary (w/ EPO)- True (long term hypoxia, RAO in eq, brachycephal syndrome in ca) or Not true (w/o hypoxia) EPO produces tumor in kidney and liver.
Polycythemia, hypovolemia
Relative- decreased plasma vol, dehydration eg. vomit, diarrhea
Polycythemia, hypervolemia
Complex- life threatening, acute stress, extreme exercise
anemia, normovolemia
absolute- several hours post acute bleeding (plasma replaces faster than cells), decreased RBC- lack of nutrients and suppression of bone marrow, decreased life span eg IHA, ectoparasitosis, sequestration of RBCs in spleen- hypersplenismus
anemia, hypovolemia
Complex- causes refusal of water, diarrhea, vomiting
anemia, hypervolemia
- Physiological- increase in plasma volume in 3rd trimester pregnancy
- Relative- increase in plasma volume, hyperhydration ie overdose fluid therapy
Serum osmolality mathematical method
2(Na+K)+urea+glucose
osmometer
Osmolality normal values
270-310mOsm/kg
Isotonic dehydration
blood, plasma loss, vomiting, diarrhea, chronic renal failure
hypertonic dehydration
primary water loss eg diarrhea, hyperventilation, chronic renal failure, ADH function loss, henle loop diuretics
hypotonic dehydration
enhanced sweating in eq, hypotonic infusion overdose: drinking lots of water after loss, hypoadrenocorticism, chronic kidney disease, diarrhea
isotonic hyperhydration
concurrent Na+ and H20 retention, overdose of IV plasma or isotonic infusion, cardiac or hepatic disorders
hypertonic hyperhydration
salt poisoning, primary and secondary hyperaldosteronism, enhanced glutocorticosteroid effect
hypotonic hyperhydration
water poisoning (excessive water intake- haematuria in calves), hypotonic infusion overdose: increased ADH function, hypothyreosis
Sodium normal value
140-150mmol/l
Potassium normal value
3.5-5.5mmol/l
Chlorine normal value
100-125mmol/l
Magnesium normal value
0.8-1.5mmol/l
Calcium normal value
2.1-3mmol/l (ionised is 45-50% of this)
Causes of hypernatremia
- Decreased water intake, polyuria, vomiting, diarrhea, hyperthermia
- Increased retention in kidneys- primary (conn) and secondary (liver disease) hyperaldosteronism
- Overdose hypertonic salt solution
- Salt poisoning
Causes of hyponatremia
- Increased water intake per os in Ru
- Overdose on hypotonic fluid
- Retention of water- renal/hepatic insufficiency
- Enhanced Na+ loss- diarrhea, hypoadrenocorticism
- Sweating (eq)
- Sequestration in body cavities
- Hyperosmolality
Hyperkalemia
increased per os intake, urinary bladder rupture, overdose K+ fluids, acute kidney failure, hypoaldosteronism, acidosis
hypokalemia
alkalosis, primary and secondary hyperaldosteronism, insulin, anorexia, long term polyuria
when do you measure chloride?
vomiting, diarrhea, polyuria/polydypsia and when there is an acid base disturbance
hyperchloremia
excessive per os intake, iv overdose, decreased excretion (hyperaldosteronism) and other cases with hypernatremia
hypochloremia
abomasal displacement, vomiting, diarrhea, sweating (eq), sequestration in body cavaties, hypoadrenocorticism
tCa is measured with what sample?
spectrophotometer with serum or heparinised plasma and ortocresolphtalein
how many forms of calcium are there in the blood plasma?
3.
47% to albumin, 13% chelated, free ionised 40%
hypocalcemia
decreased intake, hypofunction of parathyroid gland (Mg deficiency), in fe after thyroidectomy, lactating animals, seizures
hypercalcemia
increased intake, hyperfunction of parathormone, hyper A vitaminosis in fe, damages bone, soft tissue calcification
Magnesium
ATP metabolism (ATP bound to MG2+)-catalysator actin-myosin activator (maintains neuromuscular irritability) facilitates synthesis and breakdown of Ach
Magnesium indications
hypocalcemia, hypokalemia, muscle weakness, tremors, dysphagia, dyspnoa
Hypomagnesemia
decreased intake (grass tetany), reduced concentration of tMg in CSF- hyper, muscle spasms, convulsions, respiratory distress, collapse, death adults lactating are more susceptible increased excretion (renal, enteral) eg chronic diarrhea
hypermagnesemia
increased absorption after laxative agents, overdose of Mg, decreased excretion- milk fever, dehydration, addisons disease, muscle weakness, paralysis
inorganic phosphate normal amount
ca, eq- 0.8-1.8mmol/l
fe, bo, sheep- 1-2.4mmol/l
su, goat- 1-3mmol/l
inorganic phosphate
plasma buffer system, energy of RBC
in acidic environment- phophorus and ammonium molybdate- yellow phosphate molybdic acid
increases when meat/grains are fed
rate of absorption increases under Vit D effect and decreases with EPI (intestinal inflammation and Vit D deficiency)
Decreases with liver dysfunction and kidney failure
decreases with primary and secondary hyperparathyroidism
decreases with PTH (induces Ca2+ and phosphate mobilisation) and calcitonin. decreases with pH value
How to measure coagulation time
use fresh, native blood samples (2 syringe method) NO EDTA
make precise venipuncture, otherwise damaged cells will trigger tissue factor 3 and initiate the coagulation cascade.
Appearance of 1st fibrin strand- 1-2mins
CT on watch glass (treated with paraffin)- 7-15mins
CT in plastic syringe- 10-12mins
CT in glass tube- 4-5mins
CT in ACT tube (with SiO2) put in thermostat 37 degrees and check complete coag time, slowly move tube every 20-30 seconds (3 mins)
Platelet count normal amount
200-800x10^9/l
1. Burker chamber (x10^9) nor accurate
2. Blood smear (finding 1 in 1000x magnification= 20x10^9/l
finding aggregates means proper platelet function
3. ACC- particles between 5-30fl volume are taken as platelets
Clot retraction test
Blood clot gets smaller over time and serum appears around
in 1hr- serum volume of 25% of original clot size
if clot reaction is slow- thrombocytopathy
Prothrombin time factors
1,2,5,7,10,13
prothrombin normal time
10-15 seconds
how to calculate prothrombin time (PT)
within 1 hr sampling
3.8% Na2-citrate in 9:1 dilution and centrifuge for 10 mins.
separate citrated plasma from sediment (keep at 37 degrees)
reagent contains rat uterus homogenate as tissue thromboplastin (3) and CaCl2
measure time of coagulation
APTT normal time
20-30 seconds
APTT factors
1,2,5,8,9,10,11,13
how to calculate APTT
reagent contains rabbit brain homogenate as PF3 and micronised silica as contact activator
100ul citrated plasma and 100ul reagent and 100ul 0.025mmol CaCl2
measure coag time
what is dicumarol
antagonist of Vit K.
Vit K is responsible for gamma carboxylation or proconvertin (VII), Christmas (IX), Stuart Prower (X) and Prothrombin (2) as they as Ca dependent factors .
Factor VII has shortest life, prothrombin in time increases with factor VII decreasing.
Thrombin Time (TT)
citrated plasma with reagent with thombin only.
coagulation depends on fibrinogen and Factor XIII in plasma.
-Intrinsic (haemophilia A factor 8 and B factor 9 def and Von Willebrand)- Incr APTT, same PT
-Extrinsic (dicumarol, factor VII def)- same APTT, incr PT
-Common (liver dys, dicumarol, DIC, Factor X,V,II,I def)- incr APTT and PT
what is D-dimer used for
detects increased fibrinolysis (only fibrin)
very good for early detection of DIC
diagnosis of von willebrand
deficient in factor VIII
3 main parts- VW (responsible for platelet adhesion and aggregation)
VIIIc (antihaemolytic)
VIII related antigen (binds strongly to VW factor)
Dogs with this have increased BT, BMBT and decreased clot retraction ability
specific diagnosis- lack of VW antigen
methods for RBC count
- Burker chamber- anticoagulated blood with EDTA and homogenis, put 50ul blood into 9.95 phys saline and mix, add one drop into haemocytometer and count cells in 20 squares. No of counted RBC/100= RBC x10^12/l
- ACC- if particles are 40-100fl we assume they’re RBC
Normal RBC count= 4.5-8x10^12/IT/l
MCH normal size
12-30pg (younger animals bigger)
MCV normal size
60-70fl fe,eq= smaller young RBC= bigger Japanese akita has small (55-65fl) some poodles large (75-80fl)
MCHC normal size
300-350g/l
Osmotic resistance of RBC is dependent on….
pH plasma and reagents, temp, osmotic conc of plasma and reagents, RBC membrane status, regenerative status, HbF content of RBC
what is regenerative anemia
bone marrow releasing reticulocytes to make up for absence of RBC
Size of RBCs
macrocytosis, microcytosis, anisocytosis, polkylocytosis
RBC types in maturation order
Proerythroblast Basophil erythroblast Polychromatophil erythroblast Acidophil erythroblast young, mature w/o nucleus- reticulocyte
reason for appearance of young RBCs
increased production (regen anemia), spleen or bone marrow disease, leukemia, extramedullary erythrocyte production, Pb toxicosis, hyeradrenocorticism
reason for reticulocyte appearance
regen anemia, Fe deficiency anemia, haemolysis, acute and chronic blood loss
Inclusion bodies in RBCs
Heinz body- NMB- denatured Hbg
Howell-Jolly body- nuclear membrane remnants
Basophilic punctuates- nuclear membrane remnants
Is iron metabolism open or closed?
normally closed, meaning a very low amount is lost, most is reutilised.
Iron in feed (up tp duodenum)
Iron in feed is in the form Fe3+, which is not absorbable, so gastric acid frees it from these complex molecules, reducing it to Fe2+ by ascorbic acid, cysteine or glutathione in duodenum.
what happens to iron in the mucosa
it is bound to apoferritin and stored as ferritin, from here it can be absorbed to plasma or faeces.
Iron is stored in easily available form ferritin in spleen, bone marrow, liver
80-90% iron reutilised for Hgb synthesis in bone marrow
how do we lose iron?
when the ‘circle’ opens due to chronic blood loss, the iron storages deplete- iron deficiency anemia.
why do we have to use serum for iron measurement?
if we dont use serum, fibrinogen content may disturb
method for serum iron measurement
fe3+ reduced to fe2+ by ascorbic acid. it reacts with ferrosin and forms red color chelate (complex) and is measrued photometrically
*must be performed with TIBC analysis
TIBC
measure serumFe then add Fe-solution to the plasma (allows transferrin to be fully saturated, then put absorbent to solution and centrifuge (binds to free Fe sediments)
use upper layer and check seFe again.
TIBC= serum iron level (saturated)+ free transferrin (unsaturated)
causes of low TIBC
chronic inflammation, chronic liver failure, neoplastic diseases
causes of high TIBC
iron def anemia (not severe if normal iron level and high TIBC)
