BLOOD BLOOD - INTRODUCTION - PowerPoint Presentation

1. BLOOD

2. BLOOD - INTRODUCTIONPart of ECFHas two portionsCellular componentsErythrocytes (Red blood corpuscles - RBC)Leukocytes (White blood corpuscles – WBC)Thrombocytes (Platelets)Extracellular componentsWaterElectrolytesProteinsCarbohydrates (mostly glucose)LipidsEnzymesHormonesPlasma Fluid portion of blood 55-70% of whole bloodAdd anti coagulants allow cells to settle naturally / centrifugeSerumPlasma without coagulation factorsAfter formation blood clot retracts forcing clear watery serum out

3. GENERAL FUNCTIONS OF BLOODThree main functionsTransport Nutrients from digestive organs to tissueMetabolic end products from tissues to excretory organsOxygen from lungs to tissuesCarbon dioxide from tissues to lungsHormones from endocrine glands to target organs RegulationBody temperatureComposition of body fluidspH of body fluidsDefenseHumoral and cell mediated immunity Against microorganisms and other invaders

4. BLOOD CELLS - ERYTHROCYTESNon nucleated in all mammalsUsually Biconcave circular discsBiconcavity increases SA & improves gas exchangeShape and diameter varies with speciesDog – markedly biconcaveCat & horse – less biconcaveGoat – spindle shaped / fusiformCamel – ellipticalChicken RBC are nucleatedShape changes when passing through narrow vessels Volume is more important than diameter for gas exchangeMean corpuscular volume (MCV) ranges from 16 (goat ) to 65 mm3 (dog)Chicken 115 -125 mm3

5. ERYTHROCYTE CONCENTRATIONVaries greatly among speciesIntra species between individual variation is highFactors influencing RBC concentration AgeSexExerciseNutritional statusLactationPregnancyEgg productionExcitementBlood volumeStage of oestrous cycleBreed of animalTime of daySource of blood sample (venous / capillary / arterial)Environmental temperatureAltitudeAbove factors also influence haemoglobin concentrationPacked cell volumeConcentration of other blood constituents

6. SpeciesRBC in millions / mm3 or ml of bloodCattle, Pig, dog, Cat6-8Horse7-12 Sheep and goat10-14 Rabbit5.5 – 6.5 Chicken 2.5 – 3.2 CONCENTRTATION OF ERYTHROCYTES IN THE BLOOD OF DIFFERENT SPECIES(~65)(~16)(~20)(~65)(~125) (volume mm3)What is the relationship between volume and concentration of RBC?As the volume decreases the concentration of RBC increases

7. ERYTHROCYTE COMPOSITION AND FUNCTIONS62-72% of adult erythrocyte is waterRemaining solidsHemoglobin: 95% of solidsRemaining 5 % solids includeProteins of membranes & stromaLipids, Free choesterol, Neutral fatVitaminsGlucoseEnzymes – carbonic anhydrase (CA)Minerals: P, S, Cl, Mg, K , NaFunctionsTransport of hemoglobinCA helps in respiratory carbon dioxide excretionHb in RBC buffers body fluid pH

8. GENESIS OF ERYTHROCYTESEarly fetal lifeIn the yolk sacLater fetal lifeLiver, spleen & Lymph nodesLate gestation & after birthExclusively in bone marrowAdvanced ageMarrow of long bones replaced by fatSo membranous bones take over erythropoiesisWhen challenged with anemia All sites active in in fetal life could resume erythropoiesis

9. ERYTHROPIESIS IN THE ADULT Pluripotent Hemopoietic Stem Cells (PHSC) in Bone MarrowPHSCLymphoid Stem CellsCFU – BColony forming Unit BlastCFU – E(erythrocyte)ErythrocytesCFU – S(Spleen)NeutrophilsEosinophilsBasophilsMacrophagesPlateletsT-CellsB-CellsCFU – GM(Grnulocytes, Monocytes)CFU – M(Megakaryocyte)Growth Inducers:4 typesIL-3Differentiation InducersGrowth & differentiation inducers controlled by factors outside bone marrow

10. RubriblastProerythroblastBasophil ErythroblastPolychromatophil erythroblastOrthochromatophil erythroblastReticulocyte< 1% in peripheral bloodErythrocytesErythrocytesProrubricyte RubricyteMeta RubricyteReticulocyte34% Haemoglobin20m8m14-16 Erythrocytes produced from each RubriblastErythropoiesis From committed Stem Cells

11. REGULATION OF ERYTHROPOIESISRBC concentration regulated within narrow limitsAdequate for normal tissue oxygenationNot great enough to impede blood flowInadequate tissue oxygenation increases RBC in circulationAnemiaDestruction of part of bone marrow – drugs, X-raysHigh altitudes – low oxygen tension in airLevel of tissue oxygenation NOT the number of RBC in circulation

12. ERYTHROPOIETINPrincipal factor that regulates RBC productionGlycoprotein – 34KdProduced in KIDNEYS – CHIEF SOURCEMesangeal cells in glomeruliTubular epithelial cellsSmall quantity in Liver

13. Low blood volume, Anemia, Poor renal blood flow, Pulmonary disease Decreased Tissue oxygenationStimulate synthesis of ErythropoietinHaemopoietic stem cellsProerythroblastsErythrocytesRestoredTissue oxygenationROLE OF ERYTHROPOIETIN IN ERYTHROPOIESISEpinephrine, Norepinephrine, Prostaglandins

14. ROLE OF VITAMINS IN ERYTHROPOIESISCobalt in cyancobalmmine (Vit B12)Needed for maturation of erythrocytesInvolved in DNA synthesis along with Folic acidDeficiency of Vit B12 Slow maturation of RBCDefective DNA synthesisMalformation of cytoskeleton and other structuresHypochromic anemeaDeficiency of Vit. B12 and folic acidRBC large, irregular and oval in shape but not discoidMembranes become fragile – reduced life spanPyridoxine, Riboflavin, Nicotinic acid, Pantothenic acid, Biotin, thiamine and ascorbic acid also neededPyridoxine deficiency in pigs Microcytic hypochromic anemiaExcess of cobalt – polycythemia

15. LIFE SPAN AND FATE OF ERYTHROCYTESSwine – 120 daysDog – 124 daysHuman – 124 daysCow & Horse – nearer to dogChicken – 28 daysAt the end of life span RBC membrane fragilityRupture while passing through narrow capillariesTotal RBC in a 450 Kg animal = 30 trillion (30 x 1012)A v. life span of RBC = 100 daysDestruction of RBC = 0.3 trillion / dayDestruction of RBC = 35 million / secSites of RBC destructionReticuloendothelial cells of Red bone marrowBirds – Liver –Principal siteLiver also in other animals

16. HEMATOCRIT OR PACKED CELL VOLUMEHaematocritPercent of blood that is cellsPacked Cell Volume (PCV)Volume of cells per unit of bloodDetermined by centrifuging blood in haematocrit tubesNormal PCV in most species = 40% (38 - 45%)PCV = 3 X Hb concentrationFactors that change PCVHemo concentrationDehydrationAsphixiaExcitement

17. TONICITY OF RBC AND HEMOLYSISTonicity Concentration of non diffusible solutes in a solutionSuspension of RBC in hypotonic solutionsHemolysisShadow / ghost corpuscleOther causes of hemolysisFreezing and thawingStirring / agitationHigh temperatureSubstance that lower surface tensionSaponins, soaps, bile saltsAlcohol, ether, chloroform destry RBC membrane – hemolysisSuspension of RBC in hypertonic solutionsCrenationPreparation of isotonic solutions of different salts / solutes% solution = 0.03 X Mol. Wt.--------- No. ions / molecule in solutionThis formula should not be used with ammonium saltsAmmonium forms ammonia in solution, penetrates RBC & causes hemolysis

18.

19. SPECIFIC GRAVITY OF BLOODCattle : 1.052Sheep: 1.051Cat: 1.050Dog, Pig: 1.045Goat: 1.042Specif gravity of blood cells is greater than plasmaSp.gravity of RBC: 1.084 Sp.gravity of serum: 1.027Cells settle downRBC at the bottomWBC a thin layer aboveBuffy coatFinally the serum/plasma

20. ERYTHROCYTE SEDIMENTATION RATEErythrocyte sedimentation rate (ESR)Distance in mm through which upper most layer of erythrocytes pass in a unit time (usually 1h)Measured in ESR tubesEDTA is preferred anticoagulantbec. It keeps erythrocyte volume constantESR varies with speciesHorse: 15 – 38mm / 20 min (114mm/h)Ruminants: 2.4mm/7hCat: 53mm/1hPig: 1-14mm/1hChicken: 1mm/1hESR determining factor is Rouleaux formation of RBCPlasma proteins esp. fibrinogen and globulin increase Rouleaux formationCannot influence ESRSp. Gravity of RBC or plasmaSize of erythrocytesChanges in ESRMay increase with normal Hb and PCV valuesChanges are not pathognomonicMerely indicate health stautsESR increases in Acute infectionsPreseence of malignant tumorsChronic diseasesHypothyroidismpregnancy

21. HAEMOGLOBINRespiratory pigment in erythrocytesComplex iron containing conjugated proteinPigment : HemeProtein:GlobinSynthesis starts in proerythroblast continues even in reticulocytesM.Wt. 66-69Kd in diff. sp.Diff. due to globin compositionHeme is same in all animals & PlantsHb conc. 13-15g/dl in most sp.Chicken: 6.5 -9.0g/dlAdult & fetal Hb differ Fetal Hb more affinity to oxygenLess binding of 2,3-DPG to HbF

22. SYNTHESIS OF HAEMOGLOBIN2 succinyl CoA + 2 glycine Pyrrole4 Pyrrole Protoporphyrin IXProptoporphyrin IX + Fe HemeHeme+polypeptide Haemoglobin a / b2 Haemoglobin a + 2 Haemoglobin b Haemoglobin A

23. HAEMOGLOBIN AND OXYGENCombines with and dissociates from oxygen rapidlyIn the pulmonary capillariesHb combines with oxygen (oxyhaemoglobin)Oxyhaemoglobin is oxygenated product not oxide of HbR- state (relaxed)In the systemic capillariesOxyhaemoglobin looses oxygen to tissuesT-state (Taut)Oxygen carrying capacity of HbDue to iron in the heme groupOxygen binds to coordination bonds but not +ve bonds in iron Combination is loose and reversibleEasy release of oxygen to tissue as dissolved molecular oxygenAt saturation 1g of Hb carries 1.34ml of oxygen Oxyhaemoglobin is red in color (arterial blood)Reduced haemoglobin is purple in color (venous blood)

24. OTHER DERIVATIVES OF HAEMOGLOBINCarbaminohaemoglobinCombination of Hb with carbon dioxideTakes place in tissuesIn lungs Hb looses carbon dioxide and picks up oxygenCarboxy haemoglobinCombination with carbon monoxideAffinity of Hb to carbon monoxide is 20X to that of oxygen Hingh partial pressure of oxygen, carbon monoxide from carboxyhaemoglobin replacedCarboxyhaemoglobin is also red in color – misleadingMethemoglobinOxidation of ferrous iron in Hb to ferric ironTrue oxide of HbCannot act as respiratory pigmentSmall quantities formed spontaneouslyLarge quantities formed whenAminophenols, sulfonamides or oxidizing agents are givenConsumption of nitrates and chlorates also induces methemoglobin formation Common accident with farm animals consuming fertilizers / herbicidesMethaemoglobin formation induced in case of cyanide poisoning with thiosulfate

25. MYOGLOBINAlso called muscle haemoglobinHas greater affinity for oxygen than haemoglobinMyoglobin is replenished with oxygen when muscles are at restMyoglobin stores oxygen unlike haemoglobin

26. ANEMIADeficiency of Haemoglobin or RBC Due to rapid loss or slow production of RBCBlood loss anemia Due to hemorrhage Plasma volume replaced within 1-3 days But RBC replacement needs 3-4wksMicrocytic hypochromic anemia Chroninc blood loss Decreases iron absorption from GIT Reduced Hb in new RBC Pyridoxine defficiency (esp. in Pigs)Aplastic anemia Due to depression of bone marrow Too much radiation, drugs or pollutantsMegaloblastic anemia Due to deficiency of cynacobalamine and folic acidPernecious anemia Defective absorbtion of vitamins in GITHemolytic anemia Due to genetic disordersSickle cell anemiaErythroblastosis foetalis

27. LEUKOCYTES / WHITE BLOOD CORPUSCLES (WBC) Less in number than RBCFound attached to / passing through endothelial lining> 50% in marginal poolFunction in tissues RBC function in bloodDivided intoGranulocytesNeutrophilsEosinophilsBasophilsAgranulocytesLymphocytes & Plasma cellMonocytesGranulocytes and agranulocytes look after body’s defense in two different ways

28. PRODUCTION OF LEUKOCYTESFrom the committed haemopoietic stem cellsGranulocytes and monocytes formed exclusively in bone marrowLymphocytes and plasma cells from various lymphoid organsLymph glands, spleen, thymus, tonsils and lymphoid tissue in bone marrow, gut etc Pluripotent Hemopoietic Stem Cells (PHSC) in Bone MarrowPHSCLymphoid Stem CellsCFU – BColony forming Unit BlastCFU – E(erythrocyte)ErythrocytesCFU – S(Spleen)NeutrophilsEosinophilsBasophilsMacrophagesPlateletsT-CellsB-CellsCFU – GM(Grnulocytes, Monocytes)CFU – M(Megakaryocyte)

29. DEVELOPMENT STAGES IN LEUKOCYTE PRODUCTIONLymphoblastProlymphocyteLymphocyteMonoblastPromonocyteMonocyte

30.

31. LIFE SPAN OF LEUKOCYTESPresent in circulation temporarily while being transported into tissuesShort circulatory life span Granulocytes: 4-8h in circulation 4-5 days in tissuesTissue life shorter in serious infectionsMonocytes : 10-20h in circulation several months – years in tissuesLymphocytes exit and enter circulation repeatedlyLife span of lymphocytes: months to years

32. SpeciesTotal WBC / mlProportion (%) of each leukocyte typeNeutrophilsLymphocytesMonocytesEosinophilsBasophilsCow, sheep7-10k25-3060-6552-5<1Goat8-12k35-4050-5552-5<1Horse8-11k50-6030-405-62-5<1Pig15-22k30-3555-605-62-5<1Dog9-13k65-7020-255-62-5<1Cat10-15k55-6030-355-62-5<1Chicken20-30k25-3055-60103-81-4LEUKOCYTE PROFILES OF DIFFERENT DOMESTIC ANIMALS

33. NEUTROPHILSAbundant fine granular cytoplasm stains with neutral dyesNucleus of mature cells divided into lobes / segments connected by filamentsImmature cells present curved / coiled / rod like nucleus – band cellsMotile cells - diapedesis and ameboid motionActively phagocytic - Bacteria, viruses, small particlesMove to sites of inflammation through chemotaxisGranules in cytoplasm contain lysosomes Destroy phagocytosed material and neutrophilsRelease pyrogens that increase body temp.Helps other WBC to fight infectionsSlows reproduction of bacteria &virusesShift to leftIncrease in immature neutrophils - Bacterial infectionsShift to RightIncrease in hypermature neutrophils – megaloblastic anemia Usually inherited anomalyAvian BloodHeterophil is similar to neutrophilLarge rod / spindle shaped acidic granules which stain with eosin

34. CHEMOTAXISMovement of neutrophils and monocytes towards source of certain chemicalsInflamed tissues usually release these chemicalsBacterial toxins, viral toxins,degeneration products of inflammed tissue and products of plasma clotting and complementary complexChemotaxis is effective up to 100mmNo tissue site is > 30-50mm from a capillaryLarge number of leukocytes easily move to sites of inflammation

35. EOSINOPHILSLarge cells with acidic staining granules in cytoplasmHighly motile and slightly phagocyticNumbers increase inAllergic conditionsAnaphylactic shockParasitic infestationDestroy parasites byReleasing hydrolytic enzymesReactive forms of oxygen lethal to parasitesMajor basic proteinAccumulate in tissues that experienced allergic reactionEngulf antigen-antibody complexesLimit spread of local inflammations

36. BASOPHILSPresent in very small numberWater soluble cytoplasmic granulesStain with basic dyesUsually the biggest among leukocytesSome eosinophils may be as bigExhibit chemotaxisNO phagocytosisHave receptors for Ig EReleaseHeparinHistamineBradykininSerotoninLyoszomal enzymes Similar to mast cells in this property

37. MONOCYTESBig cells of lymphoid originLarge kidney shaped nucleusFaintly granular cytoplasmMove by chemotaxis to sites of inflammationIn circulation immature and nonphagocyticIn tissues become macrophages – fixed / mobileEngulf particles larger than bacteria such as dead cells~ 100 bacteria engulfed by each macrophageForm reticuloendothelial system of the bodyMacrophages could divide and produce more macrophages in tissuesMacrophages in idff. OrgansSkin – histiocytesLiver – Kupffer cellsLungs – giant cellsGiant cell limit the spread of offending agent to a local area

38. MACROPHAGE AND NEUTROPHIL RESPONSE TO INFLAMMATIONActivity starts within minutes of inflammationFirst line of defense – immediately Small number of macrophages already present in tissuesBecome mobile and phagocyticLast for ~1h2nd line of defense – within 1hNeutrophils reach inflamed area & begin phagocytosisNeutrophils in blood increase by 5X within a few hours3rd lines of defense – within 4-5 dyasNew monocytes reach the inflamed areaBetter phagocytic power and long life4th line of defense – during first 1 weekIncreased production of granulocytes and monocytes Can continue for years if properly stimulated

39. LYMPHOCYTESMost numerous among leukocytesLarge round nucleus & limited cytoplasmMotile but not phagocyticAntiviral compound – Interferon – producedTwo types of lymphocytes - Both originate in bone marrow stem cellsB-lymphocytesProcessed in Bursa fabricious in birdsProcessed and differentiated in liver, spleen and bone marrowProvide humoral immunity via antibodies against extra cellular bacteria & virusesAntigens activate B-cellsActivated B-cells form clones of plasma cells that produce Ag specific AntibodiesLife span 4-5 days T-LymphocytesProcessed and differentiated in thymusActivated by fungal , plants, neoplastic , intra cellular pathogenic antigensFrom antigen specific clones and attack the pathogens directlyBut do not produce antibodiesLife span 1-3 yrs Produce lymphokines - Inhibit / stimulate neutrophils & macrophage motilityThrough T and B cells animals build up immunity against various diseases

40. LEUKOCYTOSIS, LEUKEMIA AND LEUKOPENIALeukocytosisIncrease in number of leukocytes in circulationFactors causing leukocytosisIngestion of mealTime of dayAcute infectionsLeukemiaGeneral increase in abnormal leukocytes in circulationAbundance of myelogenous / lymphgenous stems cellsUncontrolled production of leukocytesLeukopeniaDecrease in the number of leukocytes in circulationDecrease in granulocytes - GranulocytopeniaDecrease in Lymphocytes – LyphocytopeniaUsually due to bone marrow depressionRadiationsDrugs : chloramphenicols, thiouracil, benzene, antracene etc. LEUKEMIALEUKOCYTOSISLeukopeniaNormal

41. DIFFERENTIAL LEUKOCYTE COUNT100 leukocytes in a single blood smear classifiedResult expressed as percentage usuallyCould be misleadingEx 1: Cow with traumatic reticulitisNeutrophils: 70% and Lymphocytes: 19% (Normal: N:30% & L:60%)Judgment: Neutrophilia and LymphopeniaTotal leukocyte count in the affected cow: 22000/ml (Normal:7000/ml) Number/ml in affected cow: Neutrophils: 15400; Lymphocytes: 4180Numbers/ml in healthy cow : Neutrophils: 2100; Lymphocytes: 4200Revised Judgment : ONLY NEUTROPHILIA. NO LYMPHOPENIAEx 2: Animal suffering from bone marrow depressionL: 95% and granulocytes:5%Judgment: Lyphocytosis and granulocytopeniaTotal leukocyte count: 4500/mlL:4275/ml and Granulocytes: 225/ml Revised judgment: NO LYMPHOCYTOSIS. ONLY GRANULOCYTOPENIACorrect form is number per ml of bloodNeed total leukocyte countNeutrophilsLymphocytesMonocytesEosinophilsBasophils30%65%2%2%1%Neutrophils: 15400Lymphocytes: 4180

42. SPLEEN AND CELLULAR COMPONENTS OF BLOODDuring fetal life produces RBCResumes eryhtropoiesis in chronic anemiaIn the adult produces lymphocytes and monocytesImportant reservoir of RBCReleased into circulation on increased tissue demand for oxygenDestroys aged and abnormal RBCReticuloendothelial cells of venous sinuses in spleenInvolved in formation of bile pigments & iron storageHEMOLYMPHNODESPresent in ruminants onlySimilar to spleen infunctionUndertake erythropoiesis in fetal lifeAgrnulopoiesis in adults

43. THROMBOCYTESAlso called plateletsSmall color less round /rod shaped non nucleatedAv. Diameter: 3mmLarge oval and nucleated in chickenConcentration: Mammals: 450,000 ± 150,000/ml Chicken: 25, 000 – 40, 000/mlAge, sex, Species, source of blood smaple influence count Young animals usually have lower countsLambs and calves have counts equal to adults Life span : 6 – 11 days Production Fetal life: liver, spleen and bone marrow Adults: Only bone marrow

44. GENESIS OF THROMBOCYTESMyeloid stem cellMegakaryoblastPromegakaryocyteMegakaryocyteThrombocyte

45. FUNCTIONS OF THROMBOCYTESPrevention of hemorrhage - haemostasisSpecial characters to undertake haemostasisAbility to contractContractile proteins in cytoplasmActin, myosin and thrombostheninSynthesis of enzymes and storage of calciumResidual endoplasmic reticulum and Golgi bodiesGeneration of ATPMitochondriaSynthesis of prostaglandinsNecessary enzymes presentInduce growth of vascular endothelial, smooth muscle and fibroblast cellsProduction of platelet derived growth factor (PDGF)

46. PLASMAFluid portion of bloodCellular components are suspended in plasmaYellow to color less based on diet of the animalColor mainly due to BilirubinCat, dog, sheep, goat : yellowCattle & Horse: dark color Carotenes and other pigments als0 presentComposition of plasmaWater: 91 - 92%Solids 8-9%Major Proteins: albumin, globulin, Prothrombin & FibrinogenMinor proteins: enzymes, antibodies & HormonesMost plasma proteins synthesized in liverLiver malfunction reduces plasma protein concentrationProthrombin time is a liver function testOther organic constituents: NPN, Glucose, neutral fat, phospholipids, cholesterol, Inorganic constituents: 1%Ca, Mg, K, Na, Cl, S, I, Fe, Cu, Co, Mn, Zn, Se, Mo

47. FUNCTIONS OF PLASMA PROTEINSDynamic equilibrium with tissue proteinsExchanged with tissue proteins in protein malnutritionMaintain colloid osmotic pressure (COP) of bloodOppose hydrostatic pressure of bloodPrevent excess fluid filtration into tissuesAlbumin accounts for 80% of COPHypoproteinemia causes edemaMaintain normal viscosity of bloodMaintains normal blood pressure Influence suspension stability of RBCInfluence solubility of carbohydrates, lipids & others in plasmaTransport substances bound to them Ex: hormonesRegulate acid base balance in the bodyGlobulins provide defense against infections

48. PLASMA GLOBULINSClassified into: a1, a2, b1, b2, gg globulins – immuno globulinsProduced by lymphocytes & Plasma cellsFurther divided intoIgGMost abundantPresent in blood and tissuesCrosses placenta and provides immunity to fetusIgEProduced in allergic conditionsReleases histamine from basophils and mast cellsIgAFound in secretions such as saliva, tears & milkCannot cross placentaBut secreted in collastrum & provides immunity to new bornIgDRelated to recognition of antigens by B lymphocytesAnd induces proliferation of clones of B cellsIgMNaturally occuring antibody against RBC antigens

49. PLASMA ELECTROLYTESPlasma is electrically neutralTotal conc. Of anions = Total conc. Of cationsPlasma ion concn. must be expressed in mEq./L indicates the electrically neutral nature of plasmaDescribes conc. in terms of reacting particlesFormula to convert mg/dl into mEq./LmEq./L = mg/dl x 10 x valence /atomic wt.

50. REACTION OF BLOODRefers to pH of blood plasmaMeasured very carefullyAvoid loss of blood gases esp. carbon dioxideNormal blood pH = 7.4pH of arterial blood > venous blood Why?Blood pH is maintained remarkably constantIngestion of adequate electrolytes neededLarge amounts of alkali/acid added dailyBuffered by plasma proteins, renal and respiratory systems

51. BLOOD VOLUMETo decide if blood transfusion is neededDiagnosis and classification of anemiaImportant in interpreting haematological valuesPCV, Hb, RBC/WBC count, PP conc., Values change with blood volumeMislead if hemoconcentration / dilution takes place

52. MEASUREMENT OF BLOOD VOLUMEDilution technique usedPlasma volume Evans blue (T-1824) , I*Blood volume = Plasma volume x 100/ (100 – PCV)Erythrocyte volumeP*, Fe*, Cr* usedBlood volume = RBC volume X 100/PCVTrue and venous PCVVenous PCV – volume of trapped plasmaImpossible to measure trapped plasmaTrue PCV = 96% of venous PCVBody PCVPCV in large veins & arteries > in small capillariesBody PCV is the av. Of venous and capillary PCVBody PCV = 90% of venous PCVBlood volume= plasma volume x 100 / 100 – (venous PCV) (0.96)(0.90)

53. HAEMOSTASISPrevention of blood loss when blood vessels are damagedStasis of bloodFailure of blood to move in and out of a tissue/organEven minor injury to blood vesselsRisk of heavy blood lossBecause blood flows with pressure

54. HAEMOSTATIC MECHANISMSFour different mechanismsVasoconstriction / vascular spasmPlatelet plug formationBlood coagulation /clottingFibroblast repair of damaged vessels

55. VASOCONSTRICTION / VASCULAR SPASMConstriction of a damaged blood vesselOccurs immediately after damageReduces blood flowInduced by pain and tissue damage signalsMyogenic signals from damaged smooth muscles in the blood vessel wallsSerotonin and Thromboxane A2 (TXA2) from plateletsMagnitude of spasm ∞ damage

56. PLATELET PLUG FORMATIONClumping together of plateletsCloses rent (break) in the blood vesselEffective if the rent is smallSmall rents occur 100s – 1000s of times every day Plugged by platelets effectivelyTwo types of aggregation of plateletsPrimaryPlatelets in contact with damaged endothelial layerBasement membranePlatelets swell and develop pseudopodia Adhere to one another and also to damaged surfaceSecondaryOccurs if the damage is severeDue to release of TXA2 – most powerful platelet aggregatorIrreversible aggregation

57. BLOOD COAGULATIONClotting / solidification of blood> 50 Substance in blood influence clottingSome promote & others inhibitPromoting substances – pro-coagulantsInhibiting substances – anticoagulantsAnticoagulants predominate in the bodyBut when there is damage to blood vessel(s)Procagulants override anticoagulantsBlood clottingComplex protein-protein interactionActivation of inactive procoagulants in blood

58. BLOOD CLOTTING FACTORSFACTORCOMMON NAME / SYNONYMFactor IFibrinogenFactor IIProthrombinFactor IIIThromboplastin / Tissue FactorFactor IVCalciumFactor VAccelerator globulin / Labile FactorFactor VIPostulated but does not existFactor VIIProconvertin / Serum prothrombin conversion acceleratorFactor VIIIAntihemophilic Factor A (AHF A)/ Antihemophilic globulin (AHG)Factor IXChristmas Factor / antihemophilic Factor BFactor XStuart-Prower FactorFactor XIPlasma thromboplastin Anticedent(PTA)/Antihemophilic factor CFactor XIIHageman FactorFactor XIIIFibrin Stabilizing Factor (FSF) / FibrinasePrekallikreinFletcher FactorHigh Mol .wt. KininogenFitzerald Factor / HMWK

59. FACTOR VIIIThree separate propertiesAntihemophilic property (VIII AHF)Antihemophilic factorSupport to normal platelet function (VIII VWF )(Von Williebrandt factor)Antigen detection (VIII RAG)To support blood coagulation all the three properties of factor VIII must be functional

60. BLOOD COAGULATION PATHWAYSTwo distinct but related pathwaysIntrinsic mechanismwhen blood comes into contact with a foreign surfaceExtrinsic Mechanism / Tissue Factor Pathwaywhen damage involves blood vessels and surrounding tissuesBoth cause coagulation in three stepsFormation of prothrombin activatorConversion of prothrombin to thrombinConversion of fibrinogen to fibrin

61. INTRINSIC MECHANSIMS OF BLOOD COAGULATIONDivided Into FOUR phasesContact activationActivation of Factor XFormation of ThrombinFormation of insoluble Fibrin

62. INTRINSIC MECHANISM OF BLOOD COAGULATIONI. CONTACT ACTIVATION PHASEFactor XII bound to surfacePrekallikreinKallikreinFactor XIIaHMWKFactor XIFactor XIaHMWK(exposure of Blood to collagen / basement membrane of damaged blood vessels)II. FACTOR X ACTIVATIONFactor IXFactor IXaCa++Factor XFactor XaFactor VIII,PL (Platelets),Ca++ProthrombinThrombinIII. THROMBIN FORMATIONFibrinogenFibrin monomer (soluble clot)IV. FIBRIN FORMATIONFibrin Polymer(insoluble clot)Factor XIIIFactor XIIIaCa++Ca++PL (Platelets)Factor VRATE LIMITING STEP

63. EXTRINSIC MECHANSIM OF BLOOD COAGULATIONTissue TraumaTissue FactorFactor III / ThromboplastinFactor VIIFactor VIIaFactor XFactor XaThrombinProthrombinCa++Ca++Factor V

64. TWO PATHWAYS OF BLOOD COAGULATIONEXTRINSIC MECHANISMINTRINSIC MECHANISMHMWKFactor XIIFactor XIFactor IXFactor VIIIFactor IVThromboplastinFactor VIIFactor IVFactor XFactor XFactor XaProthrombinThrombinFibrinogenFibrin

65. RELATIONSHIP BETWEEN THE TWO PATHWAYS OF COAGULATIONNot competing but complementary pathsLinked through two sets of reactionsFactor VII Factor VIIaFactor XIIaFactor IX Factor IXaFactor VIIa& TFCa++TF

66. CASCADE OF REACTIONSEach reaction in the sequence is an amplification and acceleration pointEach subsequent reaction takes place fasterMultiple actions of some the factors further amplify the cascade

67. BLOOD COAGULATION IN DIFFERENT SPECIESDifferences quantitative but not qualitativeSame basic mechanismsDifferences inQuantities of coagulation proteins involved Rate at which the coagulation takes placeCoagulation times in different speciesCat, Dog, Pig < Horse, Sheep, Cow < BirdsMay be related to platelet counts

68. FIBROBALST REPAIRRepair of damaged blood vessel wallUndertaken by fibroblastsPlatelet Derived Growth Factor (PDGF)Fibroblast Growth Factor (FGF)Angiogenic factors

69. DISSOLUTION OF BLOOD CLOTRemoval of blood clot is equally importantAs formation of the clotSo that blood flow could be restoredTwo processesClot retractionFibrinolysis

70. CLOT RETRACTIONAdequate concentration of functional platelets neededThrombostheninContractile protein in plateletsContraction of platelets in the presence of ATP & Ca++Platelets attach to more than one fibrin threadAs platelets contract they pull fibrin threads togetherThis reduces clot sizePlatelets also secrete Factor XIII (FSF)FSF increase the cross bridges among fibrin threadsThis also reduces the clot sizeSerum oozes out in the process

71.

72.

73. FIBRINOLYSISTakes place in two phasesActivation of Plasminogen to PlasminDegradation of fibrin by PlasminPlasminogenPlasminEndogenous ActivationPlsminogen Activators in vascular and tissue secretionStress, pyrogens, vasoactive drugs & anoxiaIntrinsic ActivationKalliekreinExogenous ActivationUrokinase & streptokinase(urine & hemolytic Streptococci)FibrinFibrin Degradation Products (FDP)FDP:Platelet & other cell debrisPhagocytosed by RE cells

74. CONTROL OF FIBRINOLYSISControlled by factors within the bloodInhibitors of Plasminogen activatorsAntithrombin III (AT III)[9,10,11,12,kallikrein]Inhibitors of Plasmina-2 Macroglobulin

75. PREVENTION OF CLOTTING IN NORMAL BLOOD VESSELSSeveral preventive factors in circulationEndothelial surface factorsAntithrombin action of Fibrin and AT IIIHeparina-2 macroglobulinShort half life of activated coagulation factorsRapid movement of blood

76. PREVENTION OF CLOTTING IN NORMAL BLOOD VESSELS ENDOTHELIAL SURFACTORSSmoothness of endothelium prevents contact activation phase of intrinsic pathwayGlycocalyx on endothelium repels clotting factors and plateletsThrombomodulin on endothelial surface binds thrombinSlows clotting by removal of thrombinThrmobin-thrombomodulin activates protein CProtein C inactivates Factors V and VIII

77. PREVENTION OF CLOTTING IN NORMAL BLOOD VESSELS ANTITHROMBIN ACTION OF FIBRIN AND AT IIIFibrin and AT III remove thrombin from blood85-90% of thrombin formed during clotting is adsorbed to fibrin threadsPrevents spread of thrombin and localizes availability Free thrombin if any is inactivated by AT III

78. PREVENTION OF CLOTTING IN NORMAL BLOOD VESSELS HEPARINPowerful natural anticoagulantSecreted by Basophils and mast cellsBut conc. Low and therefore not significantCombines with AT III to increase activity by 100 – 1000xHeparin – Antithrombin cofactor

79. PREVENTION OF CLOTTING IN NORMAL BLOOD VESSELS ALPHA – 2 MACROGLOBULINLarge moleculeMolecular weight : 360 kdCombines with several procoagulantsEx: Kallikrein, thrombinMakes them unavailable

80. PREVENTION OF CLOTTING IN NORMAL BLOOD VESSELS SHORT HALF LIFE OF PROCOAGULANTSActivated coagulation facotrs have short BHLTherefore destroyed before they leave the area of activationPrevents blood clotting to normal areas from injured areas

81. COAGULATION DEFECTSCongenital Common in dogsHemophilia A Deficiency of Factor VIII AHFVon Williebrandt’s diseaseDeficiency of Factor VIII VWFHemoiphilia BDeficiency of Factor IXAbnormal platelet morphologyIn cows factor XI deficiency is congenitalAcquired Bone marrow depressionAnti cancer drugs, antiinflammatory agents, aspirinDeficiency of vitamin KDecreased production of coagulation factors in liver

82. ANTICOAGULANTSUsed to prevent blood clottingFor transfusionFor analytical work HeparinNaturally occuring anticoagulant0.2mg/mlSodium citrateUsed in blood for transfusion (0.2 – 0.4%)Forms insoluble calcium citrateMay cause hypocalcemia if used in excess amountsPotassium salts should not be used May cause heart attackOxalates & FluoridesSodium, potassium, ammoniumChelating agentsEDTAHeparin and EDTA keep the size of RBC constantImportant in estimation of MCV, MCH and MCHC

83. BLOOD GROUPSHeterologous vs. homologous blood transfusionHeterologous serum clumps & lyses RBCTraditionally restricted to antigens on RBCNow it includesAntigens on WBC, platelets, tissue cells, and Hb typesLandsteiner 1900 discovered A B O systemURUD

84. BLOOD GROUPS IN ANIMALSSpeciesNo. of Blood GroupsMost Common Blood GroupSystemsNaturally occurring antibodiesCattle12A B C JAnti JSheep8B C R-OAnti RHorses8A B OAnti A (not common)Pigs15A B Anti ADogs8A B Anti A (not common)

85. BLOOD TRANSFUSIONTo restore blood volume after hemorrhageTo compensate for insufficient RBC concentrationTo prevent cardiac malfunction during surgery – preoperative transfusionIn cases of low Hb concentrationSubstitution effects – Fresh / stored bloodHemostatic effects – sufficient platelets

86. BLOOD TRANSFUSION IN ANIMALSNaturally occurring antibodies are rareLow in concentration when presentBlood group antigens occur in serum alsoCattle, sheep, PigsSo if the transfused serum has any antibodiesNeutralized by antigens in recipient serumBut do not reach RBCLocation of antigens below the rim of RBCAntibody cannot bind more than one RBCFirst transfusion without determining blood groupsAvoid transfusion of A+ Pig bloodJ+ cattle bloodR+ sheep bloodA or C+ horse bloodPresence of powerful antigens in donor blood not present in recipientInduces antibody productionAvoid transfusion of the same blood later than four days

87.

88. OTHER USES OF BLOOD GROUPS IN ANIMALSQuestionable parentageTo exclude but not confirm parentageDifferentiate monozygotic and fraternal twinsBlood groups associated with production traits and disease resistanceAdditional selection criteria