1. Introduction to Blood, Erythrocytes and Anemia Clinical Pathology Ms. Canga

2. Introduction to Blood, RBCs, and Anemia Topics for discussion: Function of blood and various components Composition – Fluid – Cellular (Specific to erythrocytes) Anemia Staining

3. The Function of Blood Classified as a _____________________ tissue – Fun Fact: Approximately ___% of animals body wt. is blood. _________________________

4. Function: Transportation Carries _______, nutrients, etc… – To every living cell in the body. – Carried by _______ in erythrocytes – Nutrients, etc. are dissolved and delivered via ____________ Carries waste products of cellular metabolism – Primary waste product is ___________ – Carried to disposal organs that excrete from body __________________&____________

5. Transportation continued Transports _______________________ – From ______________ glands to target _________ Transports _______________________ – From _______________________ in to circulation – Final destination is the _____________ as needed Transports _______________________ – To site of ______________ within _________________ – Clump together to prevent further escape.

6. Function: Defense Leukocytes (Will be covered in more detail later) – Defense from foreign invaders _______________________ Platelets (Will be covered in more detail later) – Work along side 13 clotting factors in the blood Are activated when a _______________________wall is damaged. Very complex process in which ALL factors must be activated in ____________ and __________________ in order for clot to form. Each factor depends on the _____________ factor to activate it.

7. Function: Regulation Regulatory system – _______________________

8. Regulation Acid-base balance – Regulation of blood ______. – Normal range is ________________(IDEAL = _____) Higher pH = _______________________ Lower pH = _______________________ – Required for _______________________ – Assists with neutralizing acidic waste products of cellular metabolism. Which blood is MORE alkaline? Arterial or venous? Why?

9. Regulation continued Body temperature – Regulators located in the _____________are influenced by temperature of the blood that passes over them. Most dogs and cats average 101.0 – 102.5 o F Homeostasis – Body tissue fluid is maintained as __________________ as possible. – If fluid is lost in large amounts, fluid moves from ________________________ into tissues to compensate. Leaves less plasma in bloodstream, causing _______________________. – If fluids are given or excessive fluids are present, fluid moves from ______________________ into bloodstream. Excessive fluid in bloodstream causes _______________________.

10. Composition of Blood _______________________ tissue Composed of ____________and __________ – Cellular portion is composed of: _____________– Responsible for gas exchange _____________– Responsible for defense and immunity _____________– Prevent leakage of blood from vessels – Liquid portion of whole blood is called __________ Whole Blood: Blood in cardiovascular system, OR blood that contains plasma and all other components. Plasma is ~90% water.

11. “Recipe” for an RBC __________ __________ (for the synthesis of heme-) – __________ is also important in release of iron from tissues into plasma ____________________ (formation of -globin) Essential ______________ (phospholipid bilayer) __________________: helps form RBC __________________: also helps in RBC formation ____________________: works with B12 to help form RBC.

12. Formation of RBCs Formerly known as ‘__________’ Process of formation is called _______________. – Erythropoiesis takes place in __________________ and is initiated by the cytokine called __________________ (EPO). – EPO: produced by __________, is released when kidney cells detect __________in blood. – EPO acts on __________ ______and causes it to begin undergoing __________divisions, developing into several RBC precursors.

13. Maturation Cycle of an Erythrocyte

14. Morphological Changes during Maturation Rubriblast: – Is very large, with royal blue __________. – Contains a large purple nucleus in a loose __________ pattern. Prorubricyte: – Is slightly smaller, with a __________ cytoplasm. – The __________ composing the nucleus is slightly smaller and __________ and stains __________. Rubricyte: – Is smaller still, with either a __________or slightly __________ cytoplasm as it begins to acquire the __________molecules.

15. Morphological Changes during Maturation Metarubricyte: – Cytoplasm is similar in color to __________ RBC. – Nucleus is now considered __________ (becoming more dense) and stains dark blue. – (____________________ may be seen here) Reticulocyte: – __________is “pushed out” of cell leaving behind ____________________ composed of RNA. – Cytoplasm stains slightly blue, and ____________________ may be seen if stained with Wright’s stain. (Deff-Quick is a modified Wright’s stain) – Mature RBC is seen after all ____________________ is lost from reticulocyte.

16. Morphological Changes During RBC Maturation

17. The End Result: Erythrocytes

18. Erythrocytes Mature RBC is ____________________ sac. – Contains ~ _____ % water and _____ % solids (mainly __________) Canines have __________RBCs; ~__________ in diameter – μ = a micron. One micron is one millionth of a meter (VERY TINY) Cats, horses, cows, sheep and goats are smaller; ~__________ in diameter.

20. Erythrocytes - Morphology Membrane is “____________________” Can change shape, but is not __________. _______________ shape provides more ____________________ to allow for gas exchange to take place. Disk shape allows for shorter __________ distance in and out of cell compared to a sphere. – Animals with __________ biconcave shape usually have __________ number of RBCs on average.

21. Erythrocytes & Hemoglobin Carry __________ to all tissues of the body. O 2 carried through hemoglobin, which binds to the oxygen. __________ = pigment portion – produced in __________ __________ = protein portion – Produced by __________

22. Hemoglobin - structure For every heme group, there is one __________ molecule Four __________ groups attach to each globin molecule. – This means that each hemoglobin molecule can carry __________ molecules of oxygen.

23. Hemoglobin molecule

24. Hemoglobin Types __________ hemoglobin (HbE) __________ hemoglobin (HbF) __________ hemoglobin (Hb) Each type of hemoglobin is found during different stages of development in the animal.

25. Embryonic and Fetal hemoglobin Embryonic hemoglobin (HbE): is found in early developing __________. Fetal hemoglobin (HbF): is present in fetal blood during mid to late __________. – Is also present up to a couple of months after birth. – Highest concentration of HbF is at __________.

26. Adult Hemoglobin (Hb) Found in the RBCs of all animals beginning a couple of weeks to months after birth. Gradually replaces __________ as primary type being produced by the body. Inside normal RBCs, Hb is ___________________.

27. Function of (Adult) Hemoglobin Transports __________to the tissues Exists in two normal physiological states. ____________________: Carrying oxygen – One oxygen molecule is associated with each iron molecule. – pH, temperature, and O 2 and CO 2 levels influence ability of Hb to carry O 2 ____________________: Has given up oxygen. – Also known as __________ hemoglobin. – CO2 is transported directly and indirectly in the RBC and is dissolved in the __________.

28. RBC Life Span and Destruction Average in dogs is ~____ days Average in cats is ~____ days As an RBC ages, they are replaced by young, but mature RBCs from __________ in constant ______________________ cycle. Process of aging is called ____________________.

29. Senescence The process of cellular __________ Enzyme activity __________ Cell loses ____________________ and becomes __________ as its volume decreases. 90% of destruction of senescent RBCs occurs through ____________________. – Takes place __________ the cardiovascular system via macrophage ____________________. – Macrophages of __________ are especially active in removal of senescent RBCs

30. Extravascular Hemolysis Once inside a macrophage, the __________________ of the RBC is destroyed. Contents are recovered from the macrophages and ________is transported back to red bone marrow. Heme is eliminated from the body by being converted to ____________. Bilirubin binds to ___________ (plasma protein) and is carried to the liver. Because it is not ____ soluble, it is called ________________________ or free bilirubin.

31. Extravascular Hemolysis, cont’d. Once in the liver, bilirubin is _________________ to ________________________, making the combination water soluble. Conjugated bilirubin is excreted as a ______ pigment into the intestines. Some conjugated bilirubin is converted into _____________________ by bacteria and eliminated in the ____________ as urobilin. Other conjugated bilirubin is converted into ______________________ and excreted in the __________ as stercobilin.

32. Senescence ____% of RBC destruction takes place through _________________________ hemolysis – Takes place within _____________________________. – Results in blood cell fragmentation and/or destruction When RBC membrane ruptures in blood vessel, _____ is released directly into the blood. – ____________________ Hb is picked up by transport protein called __________. – __________carries unconjugated Hb to macrophages in the __________ for breakdown. – Once in liver, ____________________hemolysis continues.

33. Senescence When ____________________is filled with ____________________ hemoglobin, the excess unconjugated Hb has nothing to bind to. Excess Hb is carried to __________ for excretion in urine. – _________________ ____hemolysis results in plasma that is pink, red, or brownish. – C/S will be ____________________. – Urine will be pink, red, or brownish in color.

34. Anemia Anemia: a pathological condition resulting in decreased oxygen-carrying capacity of the blood. May be caused by: – Low number of circulating mature RBCs Increased __________ (RBC parasites/radiation therapy) Decrease __________ (Bone marrow suppression) Inappropriate __________ (hemorrhage) – Not enough ____________________being produced for normal RBCs present. (Iron deficiency) Appropriate # of RBCs are present Insufficient Hb to fill each cell.

35. Classifying Anemia Anemia may be classified as either: – Regenerative: Usually caused by hemorrhage or hemolysis. – Non-regenerative: Involves the bone marrow (More on Anemia in next presentation)

36. Polycythemia An __________ above normal in the number of RBCs. Three common types: – _______________________________________

37. Relative Polycythemia Seen in ____________________ Common in __________ animals due to sensible and insensible losses such as: – Excessive __________, __________, __________or reduced __________intake. – You will learn more about sensible and insensible losses in Special Topics.

38. Treatment of Relative Polycythemia Correction of cause of _____________________ Use of ____________________ Managing __________ intake is critical

39. Compensatory Polycythemia Also known as __________ Polycythemia Result of __________ Bone marrow is stimulated to produce more RBCs because tissues aren’t getting enough oxygen. Animals living in ____________________ often develop this type of polycythemia. Patient in heart failure may develop this type of polycythemia because heart isn’t pumping enough blood to tissues, resulting in __________.

40. Treatment of Compensatory Polycythemia ____________________may be required if no inappropriate physiological cause If there is an underlying __________, surgical correction, followed by __________ treatment is required. (phlebotomy to normalize PCV may also be required)

41. Polycythemia Rubra Vera Rare ____________________ disorder Characterized by increased _______________ of RBCs EPO levels are __________. Bone marrow is producing __________ stimulation from EPO.

42. Treatment of Polycythemia Rubra Vera __________is required – ~__________mLs/kg Simultaneous __________therapy to maintain overall blood __________. Treatment with Rx: Hydroxyurea will help to suppress the production of RBCs.

43. Staining of Blood Different __________ within cell stain specific colors. Many different hematology stains – Different names but similar staining characteristics. – Many are _________________-_________stains (Wright’s stain, Giemsa, Leishman’s, Wright-Giemsa, and May-Grunwald are all Romanovsky-type stains) Also called ____________________ stains because they stain more than one color. Combination of basic blue and acidic red dyes dissolved in __________alcohol. Allows ____________________ structures to stain blue Allows ____________________ structures to stain red.

44. Staining Blood Stain depends on ________ you are performing. – Each test we do in here will have specific stain instructions. ____________________stain is the most widely used hematology stain. – Alkaline part is ____________________ blue – Acidic part is __________ Modified Wright’s stains offer faster staining times but don’t stain some cellular structures as effectively. – ____________________ is a Modified Wright’s stain.

45. Sources http://compepid.tuskegee.edu/syllabi/pathobio logy/pathology/clinpath/chapter2.html http://compepid.tuskegee.edu/syllabi/pathobio logy/pathology/clinpath/chapter2.html Clinical Anatomy and Physiology for Veterinary Technicians: Colville and Bassert. Second Edition Merckvetmanuals.com