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Fish Heath Management Lab 3: Hematology February 23rd. 2009 Cold Blooded Vertebrates Pokilotherms • Primary site of hematopoiesis is anterior fourth of the ___________ • Spleen is an accessory area of hematopoiesis as well as the site for blood cell destruction • Bone marrow, spleen and lymph nodes serve this function in warm blooded vertebrates • Various types of blood cells in head kidney: – – – Blood Cell Types • Hemocytoblasts- Stem Cell or blast cells – • Erythrocytes- Red Blood Cells – Carries Oxygen • Leukocytes- White Blood Cells – – – Erythrocytes • Primary Function: ____________ • Elongated elliptical cell: 13-16 m long by 7-10 wide m • Nucleus centrally located, elliptical in shape • Immature erythrocytes – less elliptical with bluish-gray cytoplasm Ghost Cells • Artifact of Sampling – Not defined – Breakdown and waste of cells • Leukocytes • ________ System Response • Coagulation • Hard to differentiate Lymphocytes • Usually sparse in blood cell smear • Spherical in shape – • Eccentrically located spherical nucleus • Neutrophils (Polymorphonuclear Leukocytes) • Only __________ leukocyte present in rainbow trout • Origin and function of theses cells not well understood • Accounts for ______of total leukocytes • Average diameter is 9-13 m ____________ nucleus (2-5 lobes) Thrombocytes • Occur in various shapes and sizes – ___________ - 5-8 m long – __________ – streaked cytoplasm on one end – ___________- streaked cytoplasm on both ends – ___________- dense nuclei, little or no cytoplasm Monocytes and Macrophages • Monocytes- – Large: 9-25 m • Macrophages: tissue dwelling monocytes – Not usually seen in blood • Monocytes are indistinguishable from _________ once in tissue • Cytoplasm often has _________ and a large nucleus Kidney Imprint • Shows various stages of hematopoiesis Blood Sampling Non-Lethal • • • Lethal • • Blood Assessment • Three things to assess: – • All leukocytes – • Hemoglobin – • What’s left (proteins, Ab, Ag, hormones, etc.) • Variations in each indices can be caused by species, sex, age, temperature, clotting, water quality, or season Hematocrit Readings • Normal – Dependent upon species – 25-40% • Low – Indicates anemia, hemorrhaging, gill damage, osmoregulation impairment – <15% • High – Indicates stress, dehydration, concentrated hemoglobin – >40% Lab Procedure • Take blood from caudal artery • Use one tube for blood smear • Seal other with CritoSeal at vacant end • Measure % of both WBCs and RBCs • Tissue imprints Tissue Imprints • Cut a small piece of Tissue • Using forceps, create imprints by touching tissue to slide in succession • Let air dry 1 minute Fish 1 K L S Stain Using Diff Quick • Fix slides in Solution A (Methyl Alcohol) for 10 minutes • Dip slides in Solution B 25 times • Dip slides in Solution C 25 times • Wash with DDI water • Dry and examine the slides