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Veterinary Hematology - An introduction Gittan Gröndahl, DVM, PhD What Is Blood, and How Much Blood Is There? B lo o d V o lu m e in A n im a ls in % o f B o d y W e ig h t Ho rse – w arm b lo o d 10 ,5 C attle – yo ung c alf 10 ,5 8 ,8 Do g 8 ,5 M o use Ham ster 8 ,2 C attle – o ld er c alf 7,3 Guinea p ig 7,2 C at 7 M o nkey 6 ,7 Rat 6 ,7 Sheep 6 ,5 Ho rse – c o ld b lo o d 6 ,5 Go at 6 ,5 C attle – c o w 6 ,5 Pig 1 2 3 5 ,6 Rab b it 5 ,4 0 2 4 6 8 10 12 3 Red Blood Cells – The Oxygen Carriers Hemoglobin Erythropoeisis: rubriblast – prorubricyte – basophilic rubricyte – polychromic rubricyte – normochromic rubricyte – metarubricyte – reticulocyte – mature erythrocyte 100 h 4 Red Blood Cells – The Oxygen Carriers Hypoxia Erythropoeitin (EPO) 5 Red Blood Cells – Measurements Hemoglobin RBC Hematocrit / PCV 6 Red Blood Cells – Physiological Alterations HGB RBC HCT growing animals... 7 Red Blood Cells – Physiological Alterations exercise, fear, stress... HGB RBC HCT 8 Red Blood Cells – Physiological Alterations dehydration, shock, high altitude, chronic lung disease, anabolic steroids... HGB RBC HCT 9 Red Blood Cells – Physiological Alterations anaemia, anaesthesia, sedation, late pregnancy... HGB RBC HCT 10 Red Blood Cells – Artefactual Alterations hemolysis, too little blood, extended storage in EDTA... HCT 11 Red Blood Cells – Regenerative Signs Reticulocytes Not normally seen in horses, cattle, sheep, goats 0-2% in dogs and swine 2-4% in rat, mouse, guinea pig, rabbit 0-5% in cats If HCT <30% (dog) or <20% (cat) count reticulocytes 12 Red Blood Cells – Regenerative Signs Reticulocytes –never in horses! Not even seen in regenerative anemia in horses Instead, macrocytes are released RDW MCV 13 Red Blood Cells – Regenerative Signs Nucleated RBC Anisocytosis greater variation in cell size, RDW Polychromasia variable coloration 14 Red Blood Cells – Regenerative Signs Howell-Jolly bodies nuclear remnants Basophilic punctuation Macrocytosis large cells, MCV 15 So – What Is This? African Gray Parrot RBC Llama RBC 16 Red Blood Cells – Number and size in mammals 17 Red Blood Cells – Mean Cell Volume (MCV) Used in classification of anemias: Macrocytosis - MCV Often in regenerative anemia Microcytosis - MCV Often sign of iron deficiency, such as chronic blood loss 18 Red Blood Cells – MCHC, Mean Cell HGB Concentration Used in classification of anemias: Normochromic - MCHC normal Hypochromic - MCHC In acute and chronic blood loss, hemolytic anemia or iron deficiency (Hyperchromic - MCHC ) Artefact! 19 Blood Groups in Animals All animals species have their specific blood group system. 1. Blood transfusions 2. Incompatibility between dam and offspring Cross-matching! 20 Anemia – Too Low Oxygen Carriage Capacity Signs of anemia: Pale in eye and mouth Vigor and strength Appetite Heart rate Respiratory rate Laboured breath Blood, bleedings, hematomas Icterus 21 Anemia – General Causes 1. Blood loss (regenerative anemia) Coagulopathies Gastrointestinal hemorrhage Platelet disorders Splenic rupture Trauma/surgery 22 Anemia – General Causes 2. Blood destruction / hemolysis (regenerative anemia) Fragmentation Immune-mediated disease Infections Intrinsic RBC defects Toxicities 23 Anemia – General Causes 3. Decreased / ineffective production of RBC (non-regenerative anemia) Anemia of inflammatory disease Aplastic or hypoplastic anemias Metabolic or endocrine disease Neoplastic disease Nutritional deficiency anemias (e.g., iron, copper, folate, cobalt) 24 Platelets – The Sealers and Healers First line of defense in damage to vessels cow Important in inflammation and wound healing cat 25 Platelets – Thrombocytopenia – PLT Caused by: Production Destruction Consumption dog Signs, if PLT <20-50 x 109/L: Hematomas Bleedings horse 26 Platelets – Thrombocytopenia – PLT Certain medications (antibiotics, NSAID, hormones) Some infections (FIV, FeLV, BVDV, EIA, Ehrlichia etc) Hemolytic anemia DIC – Disseminated intravascular coagulation Vaccination Malignant cells 27 Platelets – Thrombocytopenia – PLT False Low PLT Caused by: Platelet aggregates Platelets and fibrin clumps 28 Platelets – Thrombocytosis – PLT Caused by: Surgery or trauma Chronic bleedings Acute/chronic infections or inflammatory conditions Cushing’s disease Corticosteroid therapy Myeloproliferative disorders 29 White Blood Cells – The defence troops Defence Cleaning up Inflammatory reactions Signalling system: Cytokines Receptors 30 White Blood Cells – The defence troops First line of defence Phagocytosis Toxic proteins Parasites Allergic reactions Antibodies Cell destruction Interplay Phagocytosis Allergic reactions Histamin, heparin 31 White Blood Cells – Physiological Alterations breed, sex... WBC / 32 White Blood Cells – Physiological Alterations late pregnancy, feeding... WBC young animals... exercise, stress, excitation... 33 White Blood Cells – Artefactual Alterations extended storage... WBC Cells from very sick animals are the most sensitive 34 Granulocytes – Neutrophils White, small pink granules White/pink, no granules, ”knobby” nuclei White, no granules White, small stronger pink granules 35 Granulocytes – Band Neutrophils Bands = Left Shift Regenerative left shift Neutrophilia with >1,0 x 109/L of Bands for dogs and cats >0,3 x 109/ L of Bands for horses and cattle 36 Granulocytes – Band Neutrophils Bands = Left Shift Degenerative left shift Normal or Low WBC count with significant left shift or Band neutrophils ~ Segmented neutrophils (with any WBC count) 37 Granulocytes – Toxic Neutrophils Segmented with toxic change Segmented, normal Band with toxic change Band, normal Basophilic discoloration, foaming, Döhle bodies, toxic granules 38 Granulocytes – Eosinophils Marked variation within and between individual dogs Small rod-shaped orange granules Very large globular orange granules Many small round orange granules Gray, no granules, vacuoles 39 Granulocytes – Basophils No granules, ribbon-like nucleus, gray-lavender. Rare. Small deep purple granules. Low numbers. Small oval granules, pale lavender. Rare. Small deep purple granules. Low numbers. 40 Lymphocytes Small, dense chromatin Slightly larger Small, dense chromatin Quite variable 41 Lymphocytes - Reactive and Granular Reactive lymphocytes Associated with immune response Larger, coarse chromatin, deep blue cytoplasm Granular lymphocytes Small pink granules 42 Monocytes Extremely variable in appearance in all animals 43 Avian Hemogram 44 Dogs’ Hemogram Excitement (adrenaline = epinephrine) Stress (corticosteroids) Not so much change WBC All cells () Neutro No left shift Lymph Inflammation WBC Left shift if >1 x 109/L bands Neutro (10-30) Eos Mono 45 Cats’ Hemogram Excitement (adrenaline = epinephrine) Stress (corticosteroids) Inflammation Common reaction WBC Lymph Neutro Left shift if >1 x 109/L bands No left shift Neutro WBC (25-40) Lymph N/ Eos 46 Horses’ Hemogram Excitement (adrenaline = epinephrine) Stress (corticosteroids) Common reaction WBC (-20) WBC (12-15) No left shift Lymph (6-14) Neutro No left shift Lymph N/ Inflammation WBC Left shift if >0.3 x109/L bands Neutro (10-20) Severe infections: WBC ; Neutro ; degenerative left shift; toxic changes 47 Cattles’ Hemogram Normally: Lymph# > Neutro# so WBC DIFF is more important than WBC#! Excitement (adrenaline = epinephrine) WBC (15-27) No left shift Lymph N Stress (corticosteroids) WBC N/ No left shift Lymph Eos Inflammation Acute: WBC N/ Marked left shift (>0.3 x109/L bands) Chronic: WBC (20) Neutro Left shift 48 WBC- General Interpretation Repeated analyses -> Best for assessment of • pathological conditions • duration • prognosis 49 WBC- General Interpretation • neutrophils • mild left shift • persistent eosinophils Mild infection that the body can handle well 50 WBC- General Interpretation • neutrophils • mild left shift • lymphocytes • eosinopenia Moderate or severe infection 51 WBC- General Interpretation • immature neutrophils • segmented neutrophils Grave condition 52 WBC- General Interpretation • neutrophils • no immature neutrophils • lymphocytes • eosinophils Stress (e.g., severe disease, pain) or steroid influence 53 WBC- General Interpretation • monocytes Chronic disease 54 WBC- General Interpretation • degenerative left shift • falling lymphocyte numbers • persistent lymphopenia • persistent absence of eosinophils Each of these signs = Unfavourable prognosis 55 WBC- General Interpretation • falling WBC count together with increase in lymphocyte and eosinophil counts • decreasing numbers of immature neutrophils Each of these signs = Good prognostic signs, convalescence 56 Interpretation – Medonic histogram (dog) Platelets/Thrombocytes (PLT) Platelets and Red blood cells / Erythrocytes (RBC) White blood cells/Leukocytes (WBC) divided into 3 populations – LYM, MID and GRAN 57 Interpretation – Medonic histogram (cat) Platelets/Thrombocytes (PLT) – typical for cat: low, flat curve with poor distinction from red cells = FD-flag Platelets and Red blood cells/Erythrocytes (RBC) White blood cells / Leukocytes (WBC) divided into 3 populations – LYM, MID and GRAN 58 Interpretation – Medonic histogram (horse) Platelets/Thrombocytes (PLT) – typical for horse: low, flat curve, relatively few platelets Platelets and Red blood cells / Erythrocytes (RBC) White blood cells / Leukocytes (WBC) divided into 3 populations – LYM, MID and GRAN 59 Interpretation – Numerical values Referenc e interval PLT M PV PCT PDW HGB HCT RBC M CV M CHC RDW 281 12 0 .3 4 16 78 2 3 .4 3 .1 75 .5 332 14 .2 9 10 /L 200-500 fL 6 .1- 10 .1 H % g/L Referenc e interval L 12 0 - 18 0 3 7- 5 5 L 10 /L 5 .5 - 8 .5 L fL 6 0 - 72 H g /L 320-380 % 12 - 2 1 % 12 W BC LYM GRAN M ID 15 .3 3 .5 9 .2 2 .6 10 /L LYM % GRAN% M ID% LM L/ G/ M 23 60 17 % 9 10 /L 9 10 /L 9 10 /L Platelet count (PLT) Mean platelet volume (MPV) Plateletcrit (PCT) Referenc e interval 9 Platelet parameters 6 ,0 - 17,0 1.2 - 5 .0 Platelet distribution width (PDW) 3 .5 - 12 .0 0 .3 - 1.5 H % % 60 Interpretation – Numerical values Referenc e interval PLT M PV PCT PDW HGB HCT RBC M CV M CHC RDW 281 12 0 .3 4 16 78 2 3 .4 3 .1 75 .5 332 14 .2 9 10 /L 200-500 fL 6 .1- 10 .1 H Hemoglobin (HGB) % g/L Referenc e interval L 12 0 - 18 0 3 7- 5 5 L 10 /L 5 .5 - 8 .5 L fL 6 0 - 72 H g /L 320-380 % 12 - 2 1 % 12 W BC LYM GRAN M ID 15 .3 3 .5 9 .2 2 .6 10 /L LYM % GRAN% M ID% LM L/ G/ M 23 60 17 % 9 10 /L 9 10 /L 9 10 /L 6 ,0 - 17,0 1.2 - 5 .0 Red blood cell count (RBC) Mean cell hemoglobin concentration (MCHC = HGB/HCT) Red cell distribution width (RDW) 3 .5 - 12 .0 0 .3 - 1.5 Hematocrit (HCT = RBC x MCV) Mean red cell volume (MCV) Referenc e interval 9 Red blood cell parameters H % % 61 Interpretation – Numerical values Referenc e interval PLT M PV PCT PDW HGB HCT RBC M CV M CHC RDW 281 12 0 .3 4 16 78 2 3 .4 3 .1 75 .5 332 14 .2 9 10 /L 200-500 fL 6 .1- 10 .1 H White blood cells count (WBC) % g/L Referenc e interval L 12 0 - 18 0 3 7- 5 5 L 10 /L 5 .5 - 8 .5 L fL 6 0 - 72 H g /L 320-380 % 12 - 2 1 % 12 White blood cell parameters Lymphocytes (LYM, LYM%) Granulocytes (GRAN, GRAN%) Mid cells (MID, MID%) Referenc e interval 9 W BC LYM GRAN M ID 15 .3 3 .5 9 .2 2 .6 10 /L LYM % GRAN% M ID% LM L/ G/ M 23 60 17 % 9 10 /L 9 10 /L 9 10 /L 6 ,0 - 17,0 1.2 - 5 .0 3 .5 - 12 .0 0 .3 - 1.5 H % % 62 Why review a blood smear? To identify among the white blood cells for example…. To identify among the red blood cells for example…. Immature cells Toxic changes Monocytosis Eosinophilia Basophilia Mast cells Leukemia (blast cells) Autoagglutination Eccentrocytes Echinocytes Spherocytes Heinz bodies Nucleated red blood cells Blood parasites in or on erythrocytes Parasites in plasma, e.g. microfilaria 63 When should one review a blood smear? If you routinely make a blood smear it will always be available …… Blood samples that look very abnormal If WBC is lower or higher than normal, especially if over 30x109/L If the absolute count of lymphocytes (LYM) or mid cells (MID) is above normal If any parameter is outside normal range together with an instrument flag Blood samples with signs of anemia 64