2Reading Assignment Page 68 – Lab Pro book ‘Clinical Application’ box (Iron Deficiency Anemia) on pg. 12 of A&P bookPages 55 – 57 Lab Pro book (about counting reticulocytes)
3AnemiaLiterally means “no blood” but clinically means an ______________ ____________________below normal in any of the following values:________________________________________In other words, anemia is a condition of reduced oxygen carrying capacity of RBCsRate of RBC ______________________ = decreasedRate of RBC ______________________ = increased
4Classification of Anemia Classification is to aid in discovering the _______________ and to help guide __________________.Remember: Anemia is not a __________________, but a sign of an underlying health concern.Anemia may be considered _____________ or __________ and is generally classified in one of two different ways:Classification by RBC ________ and ____ concentrationa. RBC ____________________ (MCV, MCHC)Classification according to bone marrow response________________________________
5Aiding in Classification of Anemia A thorough ___________ must be obtained.This helps the doctor know: what the patient has been ____________ /____________, where they have been, how long they have been suffering, and possibly _________ the anemia has occurred.A physical exam should be completed.Put your ____________ on the animal!Look for _____________, _____________, _____________, active bleeding, elevated heart/respiratory rates, etc…A complete _____________evaluation is a MUST.HBC, toxins, iron deficiency, post-operative, parasites, etc….Lab tests:
6PATIENT HISTORY 1. __________________________of clinical signs ______________ onset suggests acute _________________ or ____________________________ onset suggests chronic ______________ or bone marrow depression2. Evidence of blood loss______________Blood in ______________Melena = blood in stoolHemoptysis = blood originating in lungs being expelled through nose, or coughing it up.
7PATIENT HISTORY 3. ____________________________ 4. Existence of an underlying condition or prior illness____________________________5. Exposure to drugs- human ______________ , ______________6. Exposure to toxic ______________ in the ______________- ______________ , poisonous _________, ______________
8PHYSICAL EXAMINTION ______________ Character of ___________________: Suspect: infection, leukemia, hemorrhage, or hemolysisCharacter of ___________________:______________ – liver disease or hemolysis______________ + ______________ = hemolysis______________ - hypoxia______________ or ______________ = platelet or vascular defectEcchymoses = larger than petechia.
12LABORATORY EVALUATION Initial laboratory tests to evaluate the anemic patient include (but not limited to):______________ (and color of supernatent plasma)Total ______________ proteinExamination of ______________ and ____________________________Total ______________ count______________ estimation______________ concentrationTotal ______________ count______________ **______________ evaluation **** are the two ways to classify anemia.
13PCV: Test yourself What is it measuring? Normal ranges for dogs? Normal ranges for catsPlasma (supernatent) colors?
14Plasma Protein What is it measuring? How is it measured? What is normal range for dogs and cats?
15How Many Cells should you have? As a rule, the following values should be considered:RBC total numbers should be in the ______________ .(106/μL)Plt total numbers should be in the ______________ of ______________ . (200,000 – 500,000/μL)WBC total numbers should be in the ______________ to ____________________________ . (6,000 – 17,000/μL)Neutrophils: 60 – 77%Lymphocytes: 12 – 30%Monocytes: 3 – 10%Eosinophils: 2 – 10%Basophils: rare (<2%)In dogs and cats
16Blood Film Evaluation and WBC Differential What area are you evaluating?How are cells arranged?Are RBCs normal?How many WBCs are counted?How many fields are counted for plt. estimation?What is calculation for plt. estimation?CAN look at entire slide on 10x and 40x to see how cells are appear. Following that, move to 100x and monolayer
17Total WBC CountOverall count should be in ______________ to ____________________________ . (6,000-17,000)Total count calculated by machineManual hemacytometer is rare in clinic and diluent is no longer available.Increased WBCs = ______________Decreased WBCs = ______________Diluent would lyse the RBCs and allow only WBCs to remain.
18Hemoglobin Calculation Done by machine.Aids us in calculating average ______________ of RBCs (_______)Aids us in calculating average ______ concentration within RBCs (_______)Can aid in calculating average ______________ of Hb within average RBC. (_______)*** MCH is LEAST accurate***MCH is least accurate because it uses Hb and RBC count instead of PCV.
19Classifying Anemia by RBC indices MCV: ____________________________MCHC: ____________________________MCH: ____________________________
20Rules of Thumb (ROTs): Hb concentration is ~_______ of PCV (in g/dL) Total RBCs are ~_______ of PCV (in millions)
21Classification by RBC Indicies Recall that MCV (mean corpuscular volume) describes the average volume of the individual RBCNormal MCV = _____________________Increased MCV = _____________________Decreased MCV = _____________________FORMULA: (PCV / Total RBC) X 10 = MCV (femtoliters)Normal MCV = canine: 60 – 77 fl.feline: 40 – 55 fl.
22Let’s do the math: The MCV of a patient with a PCV of 12% is: Step 1: Recall the formula:(_______/ ______________) X 10 = MCV (femtoliters)Step 2: Remember the ROTtotal RBC ≅ _______ PCV so:______________ = ______________Step 3: plug in the numbers___________________________________Is this normal for k9/fel?How would you classify this RBC?Normal for k9, not for feline. Canine = normocytic, Feline = Macrocytic
23Possible Causes of Abnormal MCV Possible causes of Increased MCV:Increased _____________________activity = #1ReticulocytosisCongenital (___________&_________________)Cats with _______ (+/- anemia)Possible causes of Decreased MCV:______________ deficiency = #1Congenital disorder (_______and ______________)Polycythemia can also cause
24Classification by RBC Indicies MCHC (mean corpuscular hemoglobin concentration):Describes the ratio of the _______of hemoglobin to the ______________in which it is contained (concentration of hemoglobin in the avg. RBC)Normal MCHC = ______________Decreased MCHC = ______________High MCHCs = artifact WHY???
25Formula (______ / ______) X ______= MCHC (g/dL) Normal MCHC = canine: ______________g/dLfeline: ______________g/dLRemember the ROTs?If you calculate MCHC by estimating Hb, the values will always come out the same.Lets do the math!
26Using the ROT The MCHC of a patient with a PCV of 33% is: Step 1: Recall the formula(_______ / _______) X _______= MCHC (g/dL)Step 2: Remember the ROTHb ≅ _______ of PCV so:_______= _______Step 3: Plug in the numbers__________________________________________
27Using actual numbers The patient’s Hb is 9g/dL, and their PCV is 30% Formula: (_______/_______) x _______SO: _______________________=_____g/dLIs this normal for k9? Fel?How would it classify the RBC?Not normal for either, both would be Hypochromic
28Low MCHC usually results from: Severe _______deficiencyMarked, regenerative anemia____________________________RBCs that do not yet have their full complement of Hb.MCHC increase:Presence of ______________, ______________, and ___________ can interfere with tests and ______________increase MCHCTrue _____________________anemia cannot exist; the erythrocyte cannot be oversaturated with ______.
29Morphologic Classification of Anemia by RBC Indicies MCHC normalMCHC decreasedMCV normalNormocyticNormochromicHypochromicMCV increasedMacrocyticMCV decreasedMicrocyticHyprochromic
31Calculating MCHYou will need to know HOW to do this for VTNE, even though it is the _______accurate of the indices.Calculates the average _______of Hb contained in average RBC.(_______/_______) x _______= MCH in picograms (pg)Normal ranges:K9: _______pgFel: _______pg
32Let’s do the Math The MCH of a patient with a PCV of 54% is: Step 1: Remember the formula(_______/_______) x _______= MCHStep 2: Remember the ROTHb ≅ _______ PCV and RBCs ≅_______ PCV_______ = _______ and _______ = _______Step 3Plug in the numbers:_____________________pg
33Classification of Anemia According to Bone Marrow Response Most pertinent of anemia classificationDistinguishes between _____________________and _____________________ anemia
34Classification Based on Bone Marrow Response Regenerative anemiaCharacterized by evidence of increased ______________ and delivery of new erythrocytes into ______________ (usually within 2-4 days).Usually suggests bone marrow is responding appropriately to either:_____________________ (acute or chronic; internal or external) or_____________________ (intravascular or extravascular)Involves determining whether absolute _____________________ numbers are increased in the blood.
35Classification According to Bone Marrow Response Nonregenerative anemiaLack of circulating ______________ RBCs in the face of _______ indicates a nonregenerative anemia and likely results from bone marrow ______________.Either reduced erythropoiesis or defective erythropoiesisNo response evident in ______________blood. (usually ______________; ______________)_____________________examination may be helpful with the diagnosis.
36Regenerative Anemia Blood Loss Anemia Acute _____________________– relatively large amount of blood lost in a brief period. (______________; ______________)PCV initially = ______________Reticulocytes should appear ~_______ hrs (peak within ~ 1 week)Causes:______________Internal or externalAccidental or surgical______________disordersBleeding ______________ or large ______________Normal PCV initially because fluid and cells lost in proportion. Takes time for spleen to release immature cells.
37Regenerative AnemiaChronic blood loss (_______Deficiency Anemia) – lost ______________and ______________for a period of time.a. Parasites______________, _______, blood-sucking _______, coccidia spp.b. GI ulcers and neoplasmsc. Inflammatory bowel diseased. Overuse of ______________donorsNote: neonates can become iron deficient due to lack of adequate dietary _______ intake.
38Iron Deficiency Anemia Body compensates for anemia by lowering _______-_______ affinity, preferential shunting of blood to vital _______, increased ______________output (tachycardia), and increased levels of _____________________.With decreasing _______ stores, erythropoiesis is limited and RBC’s become ______________and deficient in _______ (______________and _____________________).Hallmark of iron deficiency anemia is decreased _______.Keratocytes & schistocytesClinical signs include: lethargy, weakness, decrease exercise tolerance, anorexia, lack of grooming, mild systolic murmur.
39Regenerative Anemia a. Immune mediated ______________________ 2. _____________________– Increased rate of erythrocyte ______________________within the bodya. Immune mediated______________________Incompatable ______________________b. Blood parasitesHemotrophic Mycoplasmas______________________spp.
40Cytauxzoon felis inclusions Look like a ring inside the cell.
41Regenerative Anemia c. Heinz body anemia Plants Drugs or Chemicals Onions*, garlic______________________Drugs or Chemicals(______________________, Propylene glycol, Zinc, Copper, Methylene blue, Naphthalene, ______________________, phenothiazine, benzocaineDiseases (in cats)Diabetes mellitusHyperthyroidismLymphomaPropylene glycol and Ethelene glycol are both ingredients in antifreeze. Propylene = green, Ethelene = blue
42Regenerative Anemia d. ______________________ induced hemolysis RBC glycolysis is inhibited by hypophosphatemia; no glycolysis = no ATP (energy) for RBC = cell lysisDiabetic catsEnteral alimentationPhosphate = required for bone density. Also binds with ATP to create energy for the cell.
43Regenerative Anemia e. Other Microorganisms ______________________Clostridium spp. and Leptospirosis (cattle)f. ______________________intoxication (usually calves)can also occur as a result of inappropriate administration of ______________________ therapy.g. ______________________ RBC defects______________________ (shortened RBC lifespan)RBC membrane transport defectsChronic intermittent hemolytic anemia (Abyssinian and Somali cats)H2O intoxication may occur in claves given ad lib access to H20 for the first time. Excessive intake without appropriate renal response causes intravascular hemolysis due to dilution of the blood. C/s on set within 1 hour and can include: neuro, resp distress, bloat, hemoglobinuria, and death.
44Regenerative Anemia h. Miscellaneous Metabolic disorders (anything that interferes with synthesis of ______________________, RBC, etc. or anything that interferes with ______________________processes of RBC)
45Nonregenerative Anemia Most nonregenerative anemias are ______________________Further subclassified based on whether ______________________ (neutrophil production) and ______________________ (platelet production) are also affected.Animals with nonregenerative anemia in conjunction with ______________________ (neutropenia and thrombocytopenia) usually have ____________ cell injury.Possible causes: drugs, toxins, viruses (FeLV), radiation, and immune-mediated stem cell injury.
46Nonregenerative Anemia Reduced ______________________Chronic ______________________ diseaseEndocrine deficienciesInflammation and neoplasiaCytotoxic damage to the ______________________Estrogen toxicityCytotoxic cancer drug therapyChlormphenicol (cats)RadiationOther drugs
47Nonregenerative Anemia e. Infectious agentsFeLV______________________ spp.______________________f. Immune-mediatedContinued treatment with recombinant erythropoietin______________________ aplastic anemiag. Congenital/inheritedh. ______________________ and other ______________________ disorders
48Nonregenerative Anemia 2. Defective ______________________Disorders of ___________ synthesisIron, copper, and pyridoxine deficiencies; lead toxicity; drugsFolate and ____________ deficienciesAbnormal ______________________can be inherited, drug-induced or idiopathicPyridoxine = Vit B6Cobalimin = Vit B12
49Reticulocyte CountProbably the most important diagnostic tool used in the evaluation of anemia.Fewer _____________ erythrocytes are present in anemic animal; more ______________________are present.Expressed as a _____ of the RBCs present.The lifespan of a normal RBC is about 110 days (dogs) and 68 days (cats).Bone marrow should replace ___ % of the RBCs daily so the reticulocyte count should be _____-______%.
50Reticulocyte CountGently mix 4 drops of blood with 4 drops of new methylene blue in a test tube.Let mixture stand for 15 minutesUse 1 drop of mixture to prepare a diagnostic blood film and observe under high-power, oil-immersion field.Count 1,000 RBCs while separately keeping track of the number of reticulocytes (only aggregate form)Divide the reticulocyte number by 1,000 and convert to a percentage. (Multiply by 100)We will be using two blood smears and counting 500 cells on each slide to tally the 1000 cells.
51Example Reticulocyte count If you see 10 retics on slide 1 and 15 on slide 2, your total is 25 reticulocytes.25/1000 = x 100 = 2.5%
52Corrected Reticulocyte Count Performed to take in account the reduced number of circulating RBC’s in the anemic animal.Called CRC or Corrected Reticulocyte CountFORMULA:Observed retics % x PCV / normal PCV(Normal PCV: use 45% for dogs and 35% for cats)Ex: 2.5% X 30% / 45% = 1.67%Note: This calculation is necessary because the reticulocyte count is misleading in anemic patients. The problem occurs because the reticulocyte count is not really a count but rather a percentage: it reports the number of reticulocytes as a percentage of the number of red blood cells. In anemia, the patient's red blood cells are reduced, creating an elevated reticulocyte count.